Features

• Animal component-free
• Designed and optimized for small- and large-scale vaccine production
• Scalable, adaptable system to support for Commercialization
• GMP available upon request

Performance

WakoVAC Vero prototypes A and B (media 4 and 5 on the graph) have shown relatively high performances growing larger number of cells under the conditions below.

• Plate	Corning 12 well Plate　(Corning, #3513)
  Cell type	Vero cell
  Number of cells plated	30000 cells/well
  Condition	37℃, 5%CO2
  Culturing duration	5 days
  Media used	2.0 mL

• 

• 1: EMEM/5%FBS 2: DMEM/5%FBS 3: SFM from a competitor 4: WakoVAC Vero prototype A 5: WakoVAC Vero prototype B

Media Optimization Service

WakoVAC Vero will be provided as a made-to-order product, and FUJIFILM Wako is now increasing the variety of prototypes.

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Features

• Animal component-free
• Multiple prototypes are available for testing
• The latest prototype shows an improved virus yield approximately 1.5 times higher than competitor’s Medium A, commercially available serum-free media
• Designed and optimized for small- and large-scale vaccine production
• Compatible with microcarrier
• Adaptation not typically required
• Scalable and adaptable system to support for commercialization
• GMP compliant manufactured

Specification

Intended Use: For Research and Further Manufacturing

Cell culture and target expression

WakoVAC MDCK outperforms FBS contained alternative

Equal to or higher yields of influenza virus than commercially available serum-containing option

Adaptation is not necessarily required

• Switch from a commercially available serum-free medium
  
• Switch from a serum-containing medium
  

Compatible with microcarrier

Media Optimization Service

This is a made-to-order product. Please fill out your information at Media Optimization Service.

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Features

• Support wide range of insect cells
• Performs best with Sf+
  
  • Cell Growth
    

    

  • Cell Viability
    

    

  

• Fish-derived component(cGMP) and hydrolysate contained
• CoA for non-GMP and GMP available
  Cell growth, cell viability, and protein expression data can be included on CoA upon request
• Intended use: for research and further manufacturing

	Wako Cat# 160-25851	TBD
Format	Liquid	Powder
Package	1L	8kg and/or Customs upon request
Shelf-life	1 year from the date of MFG	2 years from the date of MFG
Storage Condition	2-10˚C, protect from light

Other Information

• Assure lot-to-lot consistency
• Fine protein expression as well with Sf9, Sf21 and Hi5
• L-Glutamine does not need to be supplemented
• Comparative Landscape
  
  

  Product Description	Supplier	SF	ACF	PF	CD	GMP	Hydrolysate	Cell Type	Shelf-life	Storage	Size	Bag Avaiability
  Sf Insect	FUJIFILM Irvine Scientific	Yes	Yes			Yes	+	Sf9, Sf21	1 year	2-8℃	1L	5L(MTO) Custom upon request
  WakoVAC PSFM-J1	FUJIFILM Wako	Yes	Fish-derived component	Yes		Yes	++	Sf+, Hi5, Sf9, and Sf21	1 year	2-10℃	1L	10L and 20L Powder available

Culturing Sf9 Cells

Retain robust cell growth, viability and target expression with Sf9

Cell Growth	Protein Expression Testing M: Molecular weight size marker Lane 1: Wako VAC PSFM-J1 Lane 2: Medium B Control ←: Target protein The amount of samples being used: 20μl/Lane SDS-PAGE: SuperSep™ Ace 10-20% (Wako Code: 191-15031), 200v constant voltage for 60 minutes Western blotting: PVDF membrane Antibody: Anti- his Tag antibody * Data provided by the Goshima Lab from Nagoya University
Cell Viability

Culturing Hi5 Cells

Retain robust cell growth, viability and target expression with Hi5

Cell Growth

Protein Expression Testing M: Molecular weight size marker Lane 1: Wako VAC PSFM-J1 Lane 2: Wako VAC PSFM-J1 + L-Glutamine Lane 3: Medium C Control + L-Glutamine ←: Target protein The amount of samples being used: 20μl/Lane SDS-PAGE: SuperSep™ Ace 10-20% (Wako Code: 191-15031), 200v constant voltage for 60 minutes Western blotting: PVDF membrane Antibody: Anti- his Tag antibody * Data provided by Mirai-Kougaku

Cell Viability

Target expression data with Sf21

Retain target expression with Sf21

Protein Expression Testing

• The amount of samples being used: 15μl/Lane
• SDS-PAGE: 5-20% gradient gel, 200v constant
• Voltage for 50 minutes

* Data provided by the Goshima Lab from Nagoya University

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Features

• Expressed in E. coli
• Checked residual DNA / Host cell protein
• Missed cleavage rate is lower than competitor‘s Lys-C

Specification

BSA Digestion Analysis

We incubated BSA with each protease for 1 hour and 18 hours.
After incubations, we analized the missed cleavage rate and number of peptides.

➡ The data suggested that our rLys-C have higher activity and specificity in long time reaction than the competitor’s product.

Procedure

1. Sample preparation

①Dissolve or dilute the protein sample to be digested with 25 mmol/L Tris-HCl, 1 mmol/L EDTA，pH 8.5~9.0.
②Add Disulfide threitositol (DTT) or β-mercaptoethanol to the solubilized protein at the final concentration of 5 mmol/L.
③Incubate for 30 min at room temperature.
④Add iodoacetamide to the solubilized protein at the final concentration of 10 mmol/L.
⑤Incubate in dark for 30 min at room temperature.

2. rLys-C preparation

①Dissolve lyophilized powder of rLys-C with 100 μl 12.5 mmol/L Tris-HCl, pH 8.5~9.0.

3. Sample digestion

①Add the prepared rLys-C to sample solution according to the enzyme : protein mass ratio of 1:20~1:100, and incubate at 35℃~37℃ for 2~18 hours.
②The final concentration of 0.5~1.0v/v% trifluoroacetic acid is added to stop the reaction.

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Features

• High specificity and efficiency of protein digestion allow for easy database searches by peptide mass.
• Improved cleavage at lysine residue and increase in the number of peptides are obtained by combination with trypsin.
• Packed in very small quantities according to the amounts used so that sufficient activity for in-gel digestion may be retained.

Application Data

Comparison of In-gel Digestion Using Trypsin (Tp), Lysyl Endopeptidase^® (Lep) and Lep Combined with Tp (Lep +Tp)

BSA band (100ng) resolved by SDS-PAGE was in-gel digested with Tp, Lep and Lep +Tp and analyzed by MALDI-TOFMS. The figure shows the individual mass spectra. The evaluation of these peptidases is summarized in the table.

Table: Comparison of Trypsin (Tp), Lysyl Endopeptidase^® (Lep) and Lep + Tp

Data provided by: Wada, Y., Osaka Women’s and Children’s Hospital

These results indicate there are very few missed cleavages obtained by Lep digestion. When Tp is used concomitantly with Lep, missed cleavages decrease and the number of identified peptides increase compared to when only Tp is used.

*The value resulted from subtracting the coverage obtained when database searches were performed with Missed cleavage 0 from that obtained when performed with Missed cleavage 1. “Coverage” is the percentage of peptides obtained after in-gel digestion in the whole sequence.

Mass spectrum: Comparison of Trypsin (Tp), Lysyl Endopeptidase^® (Lep) + Tp

References

1. Wada, Y. and Kadoya, M.: J. Mass Spectrom., 38, 117(2003).
2. Shevchenko, A., Wilm, M., Vorm, O. and Mann., M.: Anal. Chem., 68, 850(1996).

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beMatrix™ Gelatin

beMatrix™ Gelatin products are created using proprietary technology to yield unmatched safety and quality. Controlled for exceptionally low endotoxin levels and enhanced purity, beMatrix™ products are well suited for a range of biomedical applications including drug excipient, wound dressing materials, bone void fillers, and more.

Features

• Low endotoxin level: NMT 10 EU/g
• Virus inactivated
• IPEC-PQG GMP Compliant
• USP / EP (Ph. Eur.) / JP Compliant
• DMF / MAF registered
• Storage Condition: Room temperature
• Expiration date: 3 years from date of manufacture
• Safety Test performed
  
  

  Test Item	Cytotoxicity	Sensitization	Intradermal Reaction	Pyrogen	Antigenicity
  Result	Negative	Negative	Negative	Negative	Negative

beMatrix™ Gelatin LS-H

Features

• Alkaline-treated gelatin derived from porcine skin

• High Gel Strength (about 300g)

beMatrix™ Gelatin LS-250

Features

• Sterilized alkaline-treated gelatin derived from porcine skin
• High Gel Strength (about 250g)
• Designed for higher density and faster gelation

beMatrix™ Gelatin LS-W

Features

• Alkaline-treated gelatin derived from porcine skin
• Low Gel Strength (about 100g)

beMatrix™ Gelatin HG

Features

• Sterilized gelatin hydrolyzate derived from porcine skin
• Acute Systemic Toxicity: Negative
• Subacute Toxicity: Negative

beMatrix™ Collagen

The first collagen product used in the world’s first iPS cells based transplant.
beMatrix™ Collagen was used for culturing retinal cells derived from iPS cells that were used for treatment of “age-related macular degeneration”. Cultured cells were successfully transplant in patient without canceration, becoming the world first case.

beMatrix™ Collagen is High quality collagen with strict endotoxin control and virus inactivating processing.

beMatrix™ Collagen AT

Features

• Acid extracted collagen solution derived from porcine tendon
• Low endotoxin level: NMT 0.5 EU/mL
• Storage Condition: Cold storage [4 °C to 8 °C]
• Expiration date: 2 years from the date of manufacture
• Concentration: 3 mg/mL, pH 3

beMatrix™ Collagen TE

Features

• Pepsin-solubilized collagen solution derived from porcine skin
• Low endotoxin level: NMT 0.5 EU/mL
• Storage Condition: Cold storage [4 °C to 8 °C]
• Expiration date: 2 years from the date of manufacture
• Concentration: 5 mg/mL, pH 3

beMatrix™ Collagen FD

Features

• Freeze-dried atelocollagen derived from porcine skin
• Low endotoxin level: NMT 100 EU/g
• Storage Condition: Room temperature
• Expiration date: 2 years from the date of manufacture

Product Appearance

• Spongiform
• Size: approx. 140 x 100 x 10 mm

Application

• Scaffold
• Cell culture / transplantation
• Electro-spinning etc.

agement and virus inactivating processing.

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Antibody Information

• The paper using samples of dog¹⁾, cat²⁾, pig³⁾, marmoset⁴⁾, and zebrafish⁵⁾

References

1. Ahn, J.H., et al.: Lab. Anim. Res., 28, 3, 165 (2012).
2. Ide, T., et al.: J. Vet. Med .Sci., 72, 1, 99 (2010).
3. Gaige, S., et al.: Neurotoxicology., 34, 135(2013).
4. Rodriguez-Callejas, J.D. et al.: Front. Aging Neurosci., 8, 315(2016).
5. Fantin, A., et al.: Blood, 116, 5, 829 (2010).
6. Harlan, BA. et al.: Exp. Neurol, 327, 113219(2020).
7. Boucher, AA. et al.: J. Thromb. Haemost, 18, 91(2020).
8. Harland, M., et al.: J. Neurosci, 40, 1756(2020).
9. Ryan, KM., et al.: J. Neuroimmunol., 338, 577082(2020).
10. Jones, M. E., et al.: Brain Behav. Immun., 67, 355(2018).
11. Rui, Y., et al.: Nutrients, 10, 1084(2018).
12. Noyori, O., et al.: Clin. Transl. Immunology, 8, :e1074(2019).
13. Inyang, K. E., et al.: Pharmacol. Res., 139, 1(2019).
14. Yamamoto, M. et al.: Cell Rep., 28, 11, 2923(2019).
15. Nozaki, K., et al.: Neuropharmacology, 162, 107835(2020).
16. Sun, J.S. et al.: Mol. Med. Rep., 12, 2, 2677(2015).
17. Grishchuk, Y., et al.: Am. J. Pathol., 186, 1, 199(2016).
18. Wan, S., et al.: J. Neuroinflammation, 15, 31(2018).
19. Chen, Y., et al.: Annals of Clinical and Translational Neurology,10.1002/acn3.513 (2017).

Select Flowchart (immunostain)

References

The number of publications using FUJIFILM Wako’s Iba1 antibody per year since 2010

Searched “Iba1 019-19741 Wako” by google scholar

Application 1

Application data for immunohistochemistry, rat frozen section

Experimental condition

Code No. 019-19741/012-26723/016-26721
Tissue: Rat brain cortex
Section: Frozen section
Antibody concentration: 1/1,000

Code No. 016-26461/013-26471
Tissue: Rat brain cortex
Section: Frozen section
Antibody concentration: 1/200

These data were provided by Sanagi,T., Manabe,T., Ichinohe, N., and Kohsaka, S., National Center of Neurology and Psychiatry in Japan.

Application 2

Application Data –immunohistochemistry paraffin section-

016-27691:Anti Iba1, Rabbit(for Paraffin Section)

Experimental condition

Tissue: mouse and rat hippocampus neighborhood
Section: paraffin section
Antibody concentration: 1/1,000

Application 3

Application Data -western blotting-

016-20001: Anti Iba1, Rabbit (for Western Blotting)

Lane

1. Iba1 protein (10 ng)
2. Rat microglia (10 µg)
3. Rat neuron (10 µg)
4. Rat cortex (150 µg)

Experimental condition

SDS-PAGE: 5.5 % stacking gel, 12.5 % running gel, 100V
Blocking: 3 % skim milk in TBS at RT for 1h
Primary antibodies:anti-Iba1 antibody(1/1000) in 3 % skim milk in TTBS at 4 °C O/N
Secondary antibody: peroxidase labeled anti-rabbit antibody (1/5000) in 3 % skim milk in TTBS at RT for 1h

The data was provided by Sanagi,T., Ichinohe, N., and Kohsaka, S., National Center of Neurology and Psychiatry in Japan.

Reference of major journal

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Anti-ATRX, Monoclonal Antibody (AMab-6)

ATRX (α-thalassemia/mental retardation syndrome X-linked) is a protein involved in transcriptional regulation and chromatin remodeling. ATR-X syndrome has been known to be caused by mutation in ATRX gene on the X chromosome.
This product is a monoclonal antibody that recognizes human ATRX protein. It can be used to detect ATRX mutation, which leads to loss of ATRX protein expressionn, by immunohistochemical staining.

Application Data

Immunohistochemical staining of oligodendroglioma [A] and diffuse astrocytoma [B]

Sample: Paraffin-embedded tissue section of a lesion from a patient with brain tumor
A：Oligodendroglioma, IDH-mutant [WHO grade Ⅱ]
B：Diffuse astrocytoma, IDH-mutant [WHO grade Ⅱ]
　　Section Thickness: 4 μm
　　Antigen Activation Conditions: Solution at pH 9.0 was autoclaved for 10 minutes followed by a 40-minute storage at room temperature.
　　Quenching: 3% Hydrogen peroxide solution, 10 minutes
　　Blocking: 5% Skim milk solution, 30 minutes

Primary Antibody: 3 μg/mL Anti-ATRX monoclonal antibody (AMab-6)
Secondary Antibody: HRP-conjugated anti-mouse IgG (commercial ly-available product)
　Detection: DAB
　Counterstaining: Hematoxylin

A：Oligodendroglioma, IDH-mutant ：
　　No mutation in ATRX gene. pPositive immunohistochemical staining with anti-ATRX antibody.
B：Diffuse astrocytoma, IDH-mutant ：
　　Mutation in ATRX gene present, and loss of ATRX expression. Negative immunohistochemical staining with anti-ATRX antibody.
　　Nucleus of vascular endothelial cells (normal cells) was positively stained with anti-ATRX antibody.

Data by courtesy of Dr. Yukinari Kato at Tohoku University Graduate School of Medicine.

References

1. Ogasawara, S., Fujii, Y., Kaneko, M. K., Oki, H., Sabit, H., Nakada, M., Suzuki, H., Ichimura, K., Komori, T. and Kato, Y.： “Establishment of anti-Human ATRX monoclonal antibody AMab-6.”, Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, 35(5), 254-258 (2016).

Product Overview

・Clone No. AMab-6
・Specificity: ATRX
・Host: Mouse
・Isotype: IgG₁・κ
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (f1 μg/mL~)
 　　　　　　　　　　Immunohistochemistry (1～5 μg/mL)
 　　　　　　　　　　ELISA (1 μg/mL～)

※Please consider optimum concentrations depending on your experimental system./p>

Anti-TERT, Monoclonal Antibody (TMab-6)

Telomerase is a complex consisting of TERT (Telomerase Reverse Transcriptase), template RNAs, and regulatory subunits. TERT is the catalytic subunit of telomerase. Telomerase activity has been known to be retained not only in germ cells and tissue stem cells but also in 80%-90% of tumor cells at a high level.
This product is a monoclonal antibody against TERT, the catalytic subunit of telomerase, and is especially useful for immunohistochemical staining. The expression level of TERT protein has been known to be modified in some subtypes of glioma due to mutation in TERT promoter region. This phenomenon and aberration in ATRX gene are mutually exclusive. Mutation in TERT promoter is observed in almost all cases of glioma with IDH mutation and 1p/19q co-deletion, and also in glioma types with no IDH mutation.

Product Overview

・Clone No. TMab-6
・Specificity: TERT
・Host: Mouse
・Isotype: IgM・κ
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (1 μg/mL~)
 　　　　　　　　　　Immunohistochemistry (1 μg/mL～)
 　　　　　　　　　　ELISA (1 μg/mL～)

※Please consider optimum concentrations depending on your experimental system.

Anti-IDH, Monoclonal Antibodies

IDH (Isocitrate Dehydrogenase) is an oxidoreductase that interconverts isocitric acid and α-ketoglutarate. In humans, there are 3 types: IDH1 (cytosolic form, NADP⁺ dependent), IDH2 (mitochondrial form, NADP+ dependent), and IDH3 (mitochondrial form, NAD⁺ dependent).
IDH1IDH1 and IDH2 genes have been reported to be frequently mutated in various cancers including bile duct carcinoma, acute myeloid leukemia, chondrosarcoma, osteosarcoma, and glioma.
We provide a wide range of antibody products that recognize IDH1, IDH2, and wild-type/mutant IDH1/2.

Anti-IDH1-R132H, Monoclonal Antibody (HMab-1) [Cat. No. 018-24081]

It has been reported that mutation in IDH1 is almost limited to Arg at position 132, and R132H mutation, in which Arg at position 132 is converted to His, accounts for 80%-90%.
This product is an antibody that specifically recognizes IDH1-R132H, in which Arg at position 132 is mutated to His. It recognizes neither wild-type IDH1 nor IDH1 with non-R132H mutation.

Product Overview

・Clone No. HMab-1
・Specificity: IDH1-R132H
・Host: Mouse
・Isotype: IgG₁
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (5 μg/mL)
 　　　　　　　　　　Immunohistochemistry (5～10 μg/mL)
 　　　　　　　　　　ELISA (1～5 μg/mL)

※Please consider optimum concentrations depending on your experimental system.

References

1. Takano, S. et al.： J. Neurooncol., 108(3), 361-373 (2012).
2. Sabit, H. et al.： Brain Tumor Pathol., 31(4), 242-246 (2014).
3. Takano, S. et al.： Brain Tumor Pathol., 32(3), 169-175 (2015).
4. Kato, Y.： Brain Tumor Pathol., 32(1), 3-11 (2015).

Anti-IDH1-R132H, Monoclonal Antibody (HMab-2) [Cat. No. 013-26851]

This product, like Clone No. HMab-1, is an antibody that specifically recognizes IDH1-R132H. It recognizes neither wild-type IDH1 nor IDH1 with non-R132H mutation.
It provides a high sensitivity even at low antigen or antibody levels.

Product Overview

・Clone No. HMab-2
・Specificity: IDH1-R132H
・Host: Mouse
・Isotype: IgG₁・κ
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (1 μg/mL~)
 　　　　　　　　　　Immunohistochemistry (1 μg/mL)
 　　　　　　　　　　ELISA (1 μg/mL～)

※Please consider optimum concentrations depending on your experimental system.

References

1. 1.Fujii, Y. et al.： Biochem. Biophys. Res. Commun., 466(4), 733-739 (2015).
2. 2.Yamamichi, A. et al.： Sci. Technol. Adv. Mater., 17, 618-625 (2016).

Anti-IDH1-R132S, Monoclonal Antibody (SMab-1) [Cat. No. 015-24091]

There are IDH1 mutations in which Arg at position 132 is converted to Cys, Ser, Gly, or Leu rather than to His.
This product is a monoclonal antibody that recognizes IDH1-R132S, in which Arg at position 132 is converted to Ser.

Product Overview

・Clone No. SMab-1
・Specificity: IDH1-R132S
・Host: Mouse
・Isotype: IgG₁
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (5 μg/mL)
 　　　　　　　　　　Immunohistochemistry (5～10 μg/mL)
 　　　　　　　　　　ELISA (1～5 μg/mL)

※Please consider optimum concentrations depending on your experimental system.

References

1. Kaneko, M. K. et al.： Biochem. Biophys. Res. Commun., 406(4), 608-613 (2011).
2. Kato, Y.： Brain Tumor Pathol., 32(1), 3-11 (2015).

Anti-IDH1 monoclonal antibody (RMab-3) [Cat. No. 014-24061]

This product is an antibody that recognizes wild-type/mutant IDH1. It can be used for immunohistochemical staining of tissue sections from various types of glioma.

Product Overview

・Clone No. RMab-3
・Specificity: Wild type and mutated IDH1
・Host: Mouse
・Isotype: IgG₁
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (5 μg/mL)
 　　　　　　　　　　Immunohistochemistry (5～10 μg/mL)
 　　　　　　　　　　ELISA (1～5 μg/mL)

※Please consider optimum concentrations depending on your experimental system.

References

1. Kaneko, M. K., et al.： Tohoku J. Exp. Med., 230, 103-109 (2013).
2. Ogasawara, S. et al.： Monoclon. Antib. Immunodiagn. Immunother., 35(5), 254-258 (2016).

Anti-IDH1, Rat Monoclonal Antibody (RcMab-1) [Cat. No. 014-26381]

This product is a monoclonal antibody that recognizes wild-type/mutant IDH1. It does not recognize IDH2.
It can be used for immunohistochemical staining of brain tissue sections and western blotting of brain-derived samples as positive or loading control antibody.

Product Overview

・Clone No. RcMab-1
・Specificity: Wild type and mutated IDH1
・Host: Rat
・Isotype: IgG_2a
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (0.5 to 1 μg/mL)
 　　　　　　　　　　Immunohistochemistry (1～5 μg/mL)
 　　　　　　　　　　Immunocytochemistry(1～5 μg/mL)

※Please consider optimum concentrations depending on your experimental system./p>

References

1. Ikota, H. et al.： Brain Tumor Pathol., 32(4), 237-244 (2015).
2. Kaneko, M.K. et al.： Tohoku J. Exp. Med., 230, 103-109 (2013).
3. Kato, Y. et al.： Biochem. Biophys. Res. Commun., 432, 546-567 (2013).
4. Kaneko, M.K. et al.： Monoclon. Antib. Immunodiagn. Immunother., 32(3), 224-228 (2013).
5. Yamamichi, A. et al.： Sci. Technol. Adv. Mater., 17(1), 618-625 (2016).

Anti-IDH2, Monoclonal Antibody (RMab-22) [Cat. No. 011-24071]

This product is a monoclonal antibody that recognizes wild-type/mutant IDH2.
In mutant IDH2, Arg at position 172 is converted to Lys, Met, Gly, or Trp.
This antibody recognizes both wild-type IDH2 and IDH2-R172K/M mutant. It does not recognize IDH2-R172W.

Product Overview

・Clone No. RMab-22
・Specificity: Wild type IDH2 and mutated IDH2 (IDH2-R172K/R172M)
・Host: Mouse
・Isotype: IgG_2b
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (5 μg/mL)
 　　　　　　　　　　Immunohistochemistry (5～10 μg/mL)
 　　　　　　　　　　ELISA(1～5 μg/mL)

※Please consider optimum concentrations depending on your experimental system.

References

1. 1.Kaneko, M.K.et al.： Biochem. Biophys. Res. Commun., 432, 40-45 (2013).

Anti-mutant IDH1/2, Monoclonal Antibody (MsMab-1) [Cat. No. 015-25691]

This product is an antibody that recognizes mutated IDH1 and mutated IDH2. It does not recognize wild-type IDH1/2.
Specifically, it recognizes IDH1 mutants IDH1-R132H/R132S/R132G and IDH2 mutants IDH2-R172M/R172S/R172G.

Product Overview

・Clone No. MsMab-1
・Specificity: Mutated IDH1 (IDH1-R132H/R132S/R132G) and mutated IDH2 (IDH2-R172M/R172S/R172G)
・Host: Mouse
・Isotype: IgG_2a・κ
・Formulation: Liquid/PBS containing sodium azide
・Antibody Concentration: Approximately 1 mg/mL (stated on the product label)
・Application: Western Blot (1 μg/mL)
 　　　　　　　　　　Immunohistochemistry (5 μg/mL)
 　　　　　　　　　　ELISA(1 μg/mL)
・Reactivity: IDH1: R132S, R132G > R132H (WB)
 　　　　　IDH2　R172G > R172S > R172N (WB)

※Please consider optimum concentrations depending on your experimental system.

References

1. Kaneko, M.K. et al.： Tohoku J.Eep. Med., 230, 103-109 (2013).
2. Liu, X. et al.： Cancer Med., 2(6), 803-814 (2013).
3. Ogasawara, S. et al.： Monoclon. Antib. Immunodiagn. Immunother., 32(6), 377-381 (2013).
4. Takano, S. et al.： Brain Tumor Pathol., 32(3), 169-175 (2015).

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Product Overview

Examples of use – immunohistochemical staining-

Immunostaining of adult mouse olfactory epithelium
Green: OMP (detected with Cy2)

Data courtesy of Dr.Frank L. Margolis and Dr. Jae Hyung Koo, Department of Anatomy and Neurobiology, School of Medicine, University of Maryland

Reference

1. Keller, A., et al.: J. Neurochem., 24, 1101 (1975).
2. Baker, H., et al.: J.Comp. Neurol., 285, 246 (1989).
3. Verhaagen, J., et al.: J. Neurosci. Res., 26, 31 (1990).
4. Krishna, N. S., et al.: Brain Res., 593, 295 (1992).
5. Buiakova, O. I., et al.: Genomics, 20, 452 (1994).
6. Cummings, D. M., et al.: J. Comp. Neurol., 421, 362 (2000).
7. Koo, J. H., et al.: J. Neurochem., 90, 102 (2004).
8. Koo, J. H., et al.: J. Comp. Neurol., 487, 1 (2005).

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Anti-mouse 5-HT_1A receptor, rat monoclonal antibody (4A6)
Anti-mouse 5-HT_2C receptor, rat monoclonal antibody (6D2)

5-HT_1Areceptor and 5-HT_2C receptor are G protein-coupled receptors activated by serotonin (5-HT). Both are localized mainly in the central nervous system, and their functions controlling memory, food intake, sleep, pleasure, and anxiety, etc. Anxiolytics and antipsychotics acting on these receptors have been developed, and they are attracting attention of researchers as novel targets for drug discovery.

This product is a rat monoclonal antibody established by DNA immunization and specifically recognizes native forms of 5-HT_1A receptor and 5-HT_2C receptor.

Features

• Compatible with immunohistochemical staining
• Specifically recognizes native forms of 5-HT_1Areceptor or 5-HT_2C receptor
• Rat monoclonal antibodies established by DNA immunization

Description

DNA immunization

What is DNA immunization?

DNA immunization is a technique for generating antibodies against target proteins by using antigen proteins expressed in the animal body with expression vectors containing the target genes.

Features of DNA immunization

• Yields Antibodies specifically recognizing native forms of proteins
• Suitable for preparation of antibodies recognizing membrane proteins that were difficult to prepare by conventional methods

Principles of DNA immunization

Usage example – Immunohistochemical staining

5-HT_1Areceptor antibody

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• Localization in the cell body of prefrontal neurons was confirmed.
  This localization is consistent with the site highly expressing mRNA for 5-HT_1A.

5-HT_2C receptor

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• Localization in the cell body of prefrontal neurons was confirmed.
  This localization is consistent with the site highly expressing mRNA for 5-HT_2C.

Experimental conditions

Specimen: Prefrontal area of the brain from 10-week-old wild-type mice
Sections: 12 µm-thick frozen sections
Activation condition: Microwave treatment in a citrate buffer (pH 7.0) for 10 minutes
Antibody concentration: 1 µg/mL

Usage example – Flow cytometry

5-HT_1A receptor antibody

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• An obvious shift was observed only for cells expressing 5-HT_1A.

Anti 5-HT_2C receptor antibody

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• A clearer shift was observed for cells expressing 5-HT_2C, compared with the competitor.

Usage example – Immunohistochemical staining of the site expressing mRNA for 5-HT_1A

Positive signals were noted in amygdala (BLA), entorhinal cortex (Ent), interpeduncular nucleus (IP), and dorsal raphe nuclei (DRN).
This localization is consistent with the site highly expressing mRNA for 5-HT_1A.

Experimental conditions

Specimens: Various areas of the brain from 10-week-old wild-type mice
Sections: 12 µm-thick frozen sections
Activation condition: Microwave treatment in a citrate buffer (pH 7.0) for 10 minutes
Antibody concentration: 1 µg/mL

Usage example – Immunohistochemical staining of the site expressing mRNA for 5-HT_2C receptor

Positive signals were noted in nucleus accumbens shell (NAc shell), amygdala (BLA), and dorsal raphe nuclei (DRN).
This localization is consistent with the site highly expressing mRNA for 5-HT_2C.

Experimental conditions

Specimens: Various areas of the brain from 10-week-old wild-type mice
Sections: 12 µm-thick frozen sections
Activation condition: Microwave treatment in a citrate buffer (pH 7.0) for 10 minutes
Antibody concentration: 1 µg/mL

All immunohistochemical staining image data presented above are courtesy of Dr. Matsuda, Graduate School of Pharmaceutical Sciences, Osaka University in Japan and Dr. Takuma and Hasebe of Graduate School of Dentistry, Osaka University in Japan.

Anti-mouse serotonin transporter, rat monoclonal antibody (R5-3-2)

Serotonin transporter is a 12-transmembrane protein. It incorporates extracellular serotonin in the brain into the presynapse to modulate serotonin levels. With its reported involvement in sleep, fear, and anxiety, serotonin transporter has been investigated as a target of antidepressants.

Features

• Compatible with immunohistochemical staining
• Established by DNA immunization

Description

Usage example – Immunohistochemical staining (fluorescence staining)

Immunohistochemical staining was performed for specimens from dorsal raphe nuclei (DRN) rich in serotonin neurons where expression of serotonin transporter has been reported. As a result, fluorescence signals were noted in the neuronal cell body.

Experimental conditions

• Samples: Frozen sections of the dorsal raphe nuclei from the brain of 10-week-old ICR mice
• Staining method: Fluorescent antibody staining
• Activation condition: Microwave treatment in a citrate buffer (pH 6.0) for 10 minutes
• Antibody concentration: 1:100
• Primary antibody: This product
• Secondary antibody: Alexa 594 goat anti Rat IgG

Image data are courtesy of Dr. Takuma and Hasebe of Graduate School of Dentistry, Osaka University.

Protocol example

1. Wash the tissue section on the slide glass with phosphate-buffered saline containing 0.3 % Triton X-100^® (PBST).
2. Perform activation treatment in a citrate buffer (pH 6.0) with a microwave oven (10 minutes).
3. Bring the tissue section back to ambient temperature.
4. Wash the tissue section with 0.3 % PBST.
5. Block the tissue section with 5% goat serum-PBS (1 hour).
6. Incubate the tissue section with the primary antibody (this product) diluted 100-fold with 0.3 %PBST (4°C, overnight).
7. Wash the tissue section with 0.3 % PBST.
8. Incubate the tissue section with the secondary antibody (room temperature, 2 hours).
9. After washing with 0.3% PBST, mount the tissue section.

Triton X-100 is a registered trademark of Dow Chemical Company.

Usage example – Immunohistochemical staining (DAB staining)

Fluorescence signals were noted in the neuronal cell body.

Experimental conditions Samples: Frozen sections of the dorsal raphe nuclei from the brain of 10-week-old ICR mice Staining method: Enzyme-labeled antibody method (DAB staining) Activation condition: Microwave treatment in a citrate buffer (pH 6.0) for 10 minutes Antibody concentration: 1:100

Image data are courtesy of Dr. Takuma and Hasebe, Graduate School of Dentistry, Osaka University in Japan.

Protocol example

1. Wash the tissue section on the slide glass with phosphate-buffered saline containing 0.3 % Triton X-100^® (PBST).
2. Perform activation treatment in a citrate buffer (pH 6.0) with a microwave oven (10 minutes).
3. Bring the tissue section back to ambient temperature.
4. Wash the tissue section with 0.3 % PBST.
5. Treat the tissue section with 40% methanol containing 0.3 % H₂O₂ (30 minutes)..
6. Block the tissue section with 5% goat serum-PBS (1 hour).
7. Incubate the tissue section with the primary antibody (this product) diluted 100-fold with 0.3 %PBST (4°C, overnight).
8. Wash the tissue section with 0.3 % PBST.
9. Incubate the tissue section with the biotinylated secondary antibody (room temperature, 2 hours).
10. Wash the tissue section with 0.3 % PBST.
11. Incubate the tissue section with DAB. After dehydration treatment, mount the tissue section.

Triton X-100 is a registered trademark of Dow Chemical Company.

Usage example – Flow cytometry

Green: 293T cells
Red: 293T cells expressing mouse serotonin transporter

Antibody concentration: 0.1 µg/mL

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