GAPDH Antibody (13H12) - BSA Free

Images

 
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 ...read more
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 ...read more
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 ...read more
Immunohistochemistry-Paraffin: GAPDH Antibody (13H12) [NBP2-27103] - IHC-P detection GAPDH protein in a formalin-fixed paraffin-embedded section of normal human breast tissue using GAPDH antibody (clone 13H12) at 5 ...read more
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - Characterization of lncR492.(B) Northern blot of lncR494 using increasing amounts of loaded total. Black arrow signifies lncR492-specific signal at approximately 1400 ...read more
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - Serum from trauma patients induces intrinsic apoptosis resistance by up-regulating AAT expression in neutrophils. AAT protein expression was analyzed in neutrophils ...read more
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - WB detection of GAPDH protein (theoretical molecular weight 36 kDa) in lysates of Mouse cell lines (A) NIH 3T3 (B) RAW 264.7 using GAPDH antibody (clone 13H12) at a ...read more
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - analysis of GAPDH in HeLa and HEK 293 cells (25ug/lane) using anti-GAPDH antibody. Image from verified customer review.
Immunohistochemistry-Paraffin: GAPDH Antibody (13H12) [NBP2-27103] - IHC-P detection GAPDH protein in a formalin-fixed paraffin-embedded section of normal human stomach tissue using GAPDH antibody (clone 13H12) at 5 ...read more
Immunohistochemistry-Paraffin: GAPDH Antibody (13H12) [NBP2-27103] - IHC-P detection GAPDH protein in a formalin-fixed paraffin-embedded section of human colon tissue using GAPDH antibody (clone 13H12) at 5 ug/ml ...read more
Immunohistochemistry-Paraffin: GAPDH Antibody (13H12) [NBP2-27103] - IHC-P detection GAPDH protein in a formalin-fixed paraffin-embedded tissue section of human esophageal squamous cell carcinoma (SCC) using GAPDH ...read more
Simple Western: GAPDH Antibody (13H12) [NBP2-27103] - GAPDH/G3PDH Antibody (13H12) [NBP2-27103] - Simple Western lane view shows a specific band for GAPDH in 0.1 mg/ml of HeLa lysate. This experiment was performed ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Dr, Pm, Pm, Sh, Po, Bv, Ca, Ma, Gp, Re, SqSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-P
Clone
13H12
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml

Order Details

GAPDH Antibody (13H12) - BSA Free Summary

Immunogen
Amino acids between 275 and 325 of glyceraldehyde 3-phosphate dehydrogenase protein were used as the immunogen for this GAPDH antibody.
Localization
Cytoplasm, Nucleus
Marker
Cytosolic Marker
Predicted Species
Porcine (91%), Ferret (91%), Bovine (91%), Squirrel (91%), Guinea Pig (91%), Canine (91%), Reptile (91%). Backed by our 100% Guarantee.
Isotype
IgG1 Kappa
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • 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.
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.
Control
GAPDH Overexpression Lysate
Human Multi-tissue Tissue MicroArray (Cancer)
Reviewed Applications
Read 3 Reviews rated 5
using
NBP2-27103 in the following applications:

Publications
Read Publications using
NBP2-27103 in the following applications:

  • IHC
    1 publication
  • WB
    27 publications

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).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for GAPDH Antibody (13H12) - BSA Free

  • aging-associated gene 9 protein
  • EC 1.2.1
  • EC 1.2.1.12
  • EC 2.6.99.-
  • G3PD
  • G3PDH
  • GAPD
  • GAPDH
  • glyceraldehyde 3-phosphate dehydrogenase
  • glyceraldehyde-3-phosphate dehydrogenase
  • MGC88685
  • Peptidyl-cysteine S-nitrosylase GAPDH

Background

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

Limitations

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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Publications for GAPDH Antibody (NBP2-27103)(31)

We have publications tested in 7 confirmed species: Human, Mouse, Rat, Drosophila, Monkey, Primate, Sheep.

We have publications tested in 2 applications: IHC, WB.


Filter By Application
IHC
(1)
WB
(27)
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Human
(15)
Mouse
(7)
Rat
(3)
Drosophila
(1)
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(1)
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(2)
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Showing Publications 1 - 10 of 31. Show All 31 Publications.
Publications using NBP2-27103 Applications Species
Grotheer V, Skrynecki N, Oezel L et al. Osteogenic differentiation of human mesenchymal stromal cells and fibroblasts differs depending on tissue origin and replicative senescence Scientific reports Jun 7 2021 [PMID: 34099837] (WB, Human) WB Human
Venkatramanan, S, Ibar, C Et al. TRIP6 is required for tension at adherens junctions. J Cell Sci Mar 11 2021 [PMID: 33558314] (IHC, Mouse) IHC Mouse
Krassovka JM, Suschek CV, Prost M et al. The impact of non-toxic blue light (453 nm) on cellular antioxidative capacity, TGF-beta 1 signaling, and myofibrogenesis of human skin fibroblasts J. Photochem. Photobiol. B, Biol. Jul 6 2020 [PMID: 32659647] (WB, Human) WB Human
Modi, A;Singh, M;Gutti, G;Shanker, OR;Singh, VK;Singh, S;Singh, SK;Pradhan, S;Narayan, G; Benzothiazole derivative bearing amide moiety induces p53-mediated apoptosis in HPV16 positive cervical cancer cells Invest New Drugs Aug 20 2019 [PMID: 31432292] (WB, Human) WB Human
Srivastava, S;Battu, MB;Khan, MZ;Nandicoori, VK;Mukhopadhyay, S; Mycobacterium tuberculosis PPE2 Protein Interacts with p67phox and Inhibits Reactive Oxygen Species Production J. Immunol. Sep 1 2019 [PMID: 31375544] (WB, Mouse) WB Mouse
Eiraku, N;Chiba, N;Nakamura, T;Amir, MS;Seong, CH;Ohnishi, T;Kusuyama, J;Noguchi, K;Matsuguchi, T; BMP9 directly induces rapid GSK3-beta phosphorylation in a Wnt-independent manner through class I PI3K-Akt axis in osteoblasts FASEB J. Jul 31 2019 [PMID: 31365832]
Mishra P, Paital B, Jena S et al. Possible activation of NRF2 by Vitamin E/Curcumin against altered thyroid hormone induced oxidative stress via NF kappa B/AKT/mTOR/KEAP1 signalling in rat heart Sci Rep May 15 2019 [PMID: 31092832] (WB, Rat) WB Rat
Shwetha S, Sharma G, Raheja H et al. Interaction of miR-125b-5p with Human antigen R mRNA: Mechanism of controlling HCV replication. Virus Res. Sep 22 2018 [PMID: 30253192] (WB, Human) WB Human
van Deel ED, Octavia Y, de Waard MC et al. Exercise Training Has Contrasting Effects in Myocardial Infarction and Pressure Overload Due to Divergent Endothelial Nitric Oxide Synthase Regulation Int J Mol Sci Jul 6 2018 [PMID: 29986381]
Ibar C, Kirichenko E, Keepers B et al. Tension-dependent regulation of mammalian Hippo signaling through LIMD1 bioRxiv [PMID: 29440237] (WB, Human) WB Human
Show All 31 Publications.

Reviews for GAPDH Antibody (NBP2-27103) (3) 53

Average Rating: 5
(Based on 3 reviews)
We have 3 reviews tested in 3 species: Human, Mouse, Other.

Reviews using NBP2-27103:
Filter by Applications
WB
(3)
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(1)
Mouse
(1)
Other
(1)
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Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Mouse 01/14/2016
View

Summary

ApplicationWestern Blot
Sample TestedSplenocytes
SpeciesMouse
  5
reviewed by:
cynthia morimoto
WB Other 12/29/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 24465661
SpeciesOther
Western Blot GAPDH NBP2-27103
Enlarge
5
reviewed by:
Alex Schmidt
WB Human 10/09/2014
View

Summary

ApplicationWestern Blot
Sample TestedHeLa and HEK 293 cells
SpeciesHuman

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for GAPDH Antibody (NBP2-27103). (Showing 1 - 9 of 9 FAQs).

  1. We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
    • GAPDH is expressed in HeLa cell lines but whether or not it is the best option for a loading control antibody depends on the size of the protein you are interested in. GAPDH antibody is a good choice for low to mid MW targets, as GAPDH detects around 35 kDa.
  2. I want to know why GAPDH is used as a loading control antibody in WB techniques.
    • Our GAPDH antibody makes a good loading control because GAPDH is  a housekeeping gene essential to metabolism that is globally expressed in most tissue types and cell lines.
  3. I need a suitable loading control antibody that I can use in western blot of rat nerve extracts, which do you recommend?
    • This GAPDH antibody (NB300-221) is a good choice for a loading control antibody and has been cited in over 180 publications and used on rat lysates with good results.
  4. I'm looking for a loading control between 24kDa - 65kDa for use in simple western, we tried an actin antibody  we already had but it didn't work. What can you recommend?
    • Our GAPDH antibody NB300-221 has been validated in simple western and detects around 35 kDa so I think it would be a good choice for a simple western loading control antibody based on your size requirements.
  5. What secondary antibody should I use for visualization of the GAPDH antibody?
    • This GAPDH antibody is raised in mouse so you would want to use an anti-mouse secondary with the dye of your choice.
  6. Do you offer a smaller size for the GAPDH antibodies?
    • We offer sample sizes for many of our GAPDH loading control antibodies including NB300-221.
  7. I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
    • For homogenized tissue, beta-actin and GAPDH are both fine.
  8. 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.
    • 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.
  9. 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?
    • 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.

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Isotype Controls

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Blogs on GAPDH. Showing 1-10 of 13 blog posts - Show all blog posts.

Considerations for Quantitative Western blotting
By Jamshed Arslan, Pharm. D., PhD. Since its inception in 1979, Western blotting has undergone several developments. The use of radioactive probes was common throughout 1980s, but utilizing secondary antibodies labell...  Read full blog post.

The use of Beta Actin (AC-15) as a loading control across multiple species
Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two...  Read full blog post.

Tips on choosing an ideal loading control antibody for Western Blotting
Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. When p...  Read full blog post.

GAPDH - A "housekeeping" gene with diverse functions in cellular homeostasis
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a well-known housekeeping gene with functions in glycolysis. Many biologists are familiar with the gene and use GAPDH antibodies for a loading control when performing western blots. However, this ...  Read full blog post.

A New Standard in Antibody Testing - Simple Western Certified Antibodies
The Western blot is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. The tradi...  Read full blog post.

GAPDH: More than a housekeeping gene
GAPDH is a 146kD tetramer glycolytic pathway metabolic enzyme composed of four 30-40 kDa subunits. It is responsible for reversibly phosphorylating its substrate glyceraldehyde 3-phosphate within the glycolytic pathway.  Apart from its role in gl...  Read full blog post.

GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase)
GAPDH is a 146 kD tetramer glycolytic pathway metabolic enzyme responsible for reversibly phosphorylating glyceraldehyde 3-phosphate. It may have other possible functions in transcriptional activation. GAPDH is highly expressed due to this housekeepin...  Read full blog post.

Time to Start Actin Like a Reliable 'Housekeeper'!
A growing body of data and studies using actin antibodies supports a view of the actin cytoskeleton of smooth muscle cells as a dynamic structure that plays an integral role in regulating the development of mechanical tension and the material properti...  Read full blog post.

Beta Actin and GAPDH: The Importance of Loading Controls
Western blotting is an essential technique to probe protein expression in complex cell or tissue lysates. To accurately determine protein expression and interpret Western blot results, it is important to use loading controls. A loading control anti...  Read full blog post.

The GAPDH Antibody in Western blot Assays
The loading controls on our antibody database are widely used in gel electrophoresis and Western blotting studies. Products like the GAPDH antibody detect "housekeeping" proteins which are abundantly distributed in cells. This makes them useful for ch...  Read full blog post.

Showing 1-10 of 13 blog posts - Show all blog posts.
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Recent Reviews

5
5
3
4
0
3
0
2
0
1
0

 
Verified Customer
01/14/2016
Application: WB
Species: Mouse

 
cynthia morimoto
12/29/2014
Application: WB
Species: Other

Alex Schmidt
10/09/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot