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Key Product Details

Species Reactivity

Human, Mouse, Rat, Drosophila, Monkey, Primate, Sheep

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 13H12

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for GAPDH Antibody (13H12) - BSA Free

Immunogen

Amino acids between 275 and 325 of glyceraldehyde 3-phosphate dehydrogenase protein were used as the immunogen for this GAPDH antibody.

Predicted Species

Bovine (91%), Canine (91%), Ferret (91%), Guinea Pig (91%), Porcine (91%), Reptile (91%), Squirrel (91%). Backed by our 100% Guarantee.

Reactivity Notes

Based upon 91% sequence similarity with immunogen, this antibody is predicted to react with Guinea Pig, Sheep, Squirrel, Porcine/Pig, Ferret, Canine/Dog/Cat, Bovine, Reptile / Rattlesnake and several other species. Immunogen shows 82% similarity to Xenopus and Zebrafish. Rat, sheep, and monkey reactivity reported in scientific literature (PMID: 24796753, PMID: 27618403, and PMID: 24462973 respectively).

Localization

Cytoplasm, Nucleus

Marker

Cytosolic Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

36 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for GAPDH Antibody (13H12) - BSA Free

Immunocytochemistry/Immunofluorescence: GAPDH Antibody (13H12) [NBP2-27103] - GAPDH antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). A dilution of 1:10 was used. Image objective 40x.
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - WB detection of GAPDH protein (theoretical molecular weight: 36 kDa) in HeLa cells lysate using GAPDH antibody (clone 13H12) in (A) the absence and (B) the presence of immunizing peptide.
Immunohistochemistry-Paraffin: GAPDH Antibody (13H12) [NBP2-27103] - IHC-P detection GAPDH protein in a formalin-fixed paraffin-embedded section of human rectal carcinoma tissue using GAPDH antibody (clone 13H12) at 5 ug/ml concentration.

Applications for GAPDH Antibody (13H12) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:10

Immunohistochemistry

5 ug/ml

Immunohistochemistry-Paraffin

5 ug/ml

Simple Western

1:25

Western Blot

0.25 - 1 ug/ml
Application Notes
GAPDH is a widely used loading control for quantitative Western blotting. In IHC-P, the staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. WB reported in a verified customer review.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 5 using NBP2-27103 in the following applications:

Published Applications

Read 32 publications using NBP2-27103 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: GAPDH

Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis, converting glyceraldehyde-3-phosphate into 1,3 diphosphoglycerate. This constitutively expressed, homotetramer protein can be found in the nucleus and cytoplasm, and the monomer has a theoretical molecular weight of 36 kDa. Known as a housekeeping gene, GAPDH routinely serves as a control for real-time PCR (RT-PCR) and as a loading control for Western Blots (1-3). In addition to its role in glycolysis, GAPDH has multiple cellular functions including membrane trafficking, transcription activation, apoptosis, DNA repair, and has been implicated in various pathologies such as cancer and neurodegenerative diseases including Alzheimer's and Huntington's (4,5).

References

1) Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3):389-95. PMID: 15769908

2) Jia Y, Takimoto K. (2006) Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res. 98(3):386-93. PMID: 16385079

3) Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew TL, Green KJ. (2005) Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol. 171(6):1045-59. PMID: 16365169

4) Sirover MA1. (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1432(2): 159-84. PMID: 10407139

5) Tristan C, Shahani N, Sedlak TW, Sawa A. (2011) The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal. 23(2):317-23. PMID: 20727968

Long Name

Glyceraldehyde-3-phosphate Dehydrogenase

Alternate Names

G3PDH

Entrez Gene IDs

2597 (Human)

Gene Symbol

GAPDH

UniProt

Product Documents for GAPDH Antibody (13H12) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for GAPDH Antibody (13H12) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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FAQs for GAPDH Antibody (13H12) - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: I have GAPDH Antibody (NBP2-27103). According to the datasheet it is monoclonal. I want to do IF staining. Can it be stained with goat anti-rabbit IgG-FITC.

    A: Our GAPDH antibody with catalogue number NBP2-27103 has been validated for immunofluorescence. Our own immunofluorescence testing data was generated in HeLa cells; detection was with a Dylight 488-conjugated anti-mouse secondary antibody. Since NBP2-27103 is a mouse antibody you will need to use an anti-mouse secondary antibody for detection; a goat anti-rabbit antibody will not be suitable. NBP2-27103 is of the isotype IgG1 kappa, and you can see our anti-mouse IgG1 secondary antibodies at the following link: anti-mouse IgG1 secondary antibodies You can use the search filters at the left hand side of our webpage to select your conjugate of choice.

  • Q: I have a question about NBP2-27103 GAPDH ab for Simple Western Wes system. In the application note "In Simple Western only 10 - 15 uL of the recommended dilution is used per data point." What do you mean 10-15 micro L of dilution?

    A: It means to use only 10-15 ul of the diluted stock. Dilute from 0.25- 1 ug/ml then use 10- 15 ul on the column.

Showing  1 - 2 of 2 FAQs Showing All
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