GAPDH Antibody (NB615)

Images

 
Western Blot: GAPDH Antibody (NB615) [NB300-285] - Analysis of 40ug HeLa whole cell lysates using NB300-285 at a dilution of 1:100.
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody (NB615) [NB300-285] - IF Confocal analysis of Hela cells using GAPDH antibody (NB615) (NB300-285, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as ...read more
Western Blot: GAPDH Antibody (NB615) [NB300-285] - Western blot analysis of HeLa, Jurkat, and COS lysate using GAPDH (NB300-285) antibody at 1:100.
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody (NB615) [NB300-285] - Confocal immunofluorescence of HeLa cells using GAPDH antibody at 1:10 (green). Nuclei were counterstained with DAPI (blue).
Simple Western: GAPDH Antibody (NB615) [NB300-285] - Simple Western lane view shows a specific band for GAPDH in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Ca, Ch, RbSpecies Glossary
Applications WB, Simple Western, ICC/IF
Clone
NB615
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Order Details

GAPDH Antibody (NB615) Summary

Immunogen
Full length native GAPDH from human erythrocytes. [UniProt# P04406]
Localization
Cytoplasmic
Marker
Cytosolic Marker
Isotype
IgM
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:2000
  • Simple Western 1:50-1:300
  • Immunocytochemistry/Immunofluorescence 1:100-1:500
Application Notes
This GAPDH antibody is useful for Western blot and Immunocytochemistry/Immunofluorescence. In Western blot a band is observed at approx. 36 kDa (predicted molecular weight: 40.2 kDa). In ICC/IF, cytoplasmic staining was observed in HeLa cells.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
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 Lysate (NBL1-10967)
Reviewed Applications
Read 3 Reviews rated 4.7
using
NB300-285 in the following applications:

Publications
Read Publications using
NB300-285 in the following applications:

  • WB
    8 publications

Reactivity Notes

Bovine, canine, chicken, human, mouse, porcine, rabbit, and rat.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
10mM Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for GAPDH Antibody (NB615)

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

Background

GAPDH (glyceraldehyde-3-phosphate dehydrogenase or GAPD) is a key enzyme in glycolytic pathway, wherein it catalyzes the first step by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. It localizes mainly in the cytoplasm from where it translocates to nucleus following S-nitrosylation and interaction with SIAH1. Nuclear GAPDH implicates in transcription, RNA transport, DNA replication and apoptosis, via its nitrosylase activity which mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2, PRKDC etc. GAPDH also regulates the organization/assembly of cytoskeleton and facilitates CHP1-dependent microtubule - membrane associations. It is a component of GAIT (gamma interferon-activated inhibitor of translation) complex which mediates IFN-gamma-induced transcript-selective translation inhibition in inflammation. Because of its expression as housekeeping protein in most cell types, GAPDH is often used as a control molecule in various genes expression studies, however, recent evidence has shown the association of its altered expression with neurodegenerative pathologies such as Huntington disease, Alzheimer's disease etc., and elevated GAPDH mRNA/protein expression levels have been seen in pancreatic, lung and prostate cancers.

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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NB300-285
Species: Hu, Mu, Rt, Po, Bv, Ca, Ch, Rb
Applications: WB, Simple Western, ICC/IF

Publications for GAPDH Antibody (NB300-285)(10)

We have publications tested in 3 confirmed species: Human, Mouse, Canine.

We have publications tested in 1 application: WB.


Filter By Application
WB
(8)
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Filter By Species
Human
(5)
Mouse
(2)
Canine
(1)
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Showing Publications 1 - 10 of 10.
Publications using NB300-285 Applications Species
Kechele DO, Blue RE, Zwarycz B et al. Orphan Gpr182 suppresses ERK-mediated intestinal proliferation during regeneration and adenoma formation J. Clin. Invest Jan 17 2017 [PMID: 28094771] (WB, Mouse) WB Mouse
Shirato K, Ujike M, Kawase M, Matsuyama S. Identification of CCL2, RARRES2 and EFNB2 as host cell factors that influence the multistep replication of respiratory syncytial virus. Virus Res. 2015 Aug 12 [PMID: 26277777] (WB, Human) WB Human
Gu Y, Bouwman P, Greco D et al. Suppression of BRCA1 sensitizes cells to proteasome inhibitors. Cell Death Dis 2014 Dec 18 [PMID: 25522274] (WB) WB
Rajala HL, Eldfors S, Kuusanmaki H et al. Discovery of somatic STAT5b mutations in large granular lymphocytic leukemia. Blood 2013 Apr 17 [PMID: 23596048]
Ichiki T, Boerrigter G, Huntley BK et al. Differential Expression of the Pro-Natriuretic Peptide Convertases Corin and Furin in Experimental Heart Failure and Atrial Fibrosis Am J Physiol Regul Integr Comp Physiol 2012 Nov 14 [PMID: 23152112] (WB, Canine, Human) WB Canine, Human
Chiappinelli KB, Haynes BC, Brent MR et al. Reduced DICER1 elicits an interferon response in endometrial cancer cells. Mol Cancer Res. 2012 Mar [PMID: 22252463] (WB, Human) WB Human
Shen, L et al. Drug sensitivity prediction by CpG island methylation profile in the NCI-60 cancer cell line panel. Cancer Res;67(23):11335-43. 2007 Dec 1. [PMID: 18056460]
George, J et al. Restoration of tissue factor pathway inhibitor-2 in a human glioblastoma cell line triggers caspase-mediated pathway and apoptosis. Clin Cancer Res;13(12):3507-17. 2007 Jun 15. [PMID: 17575213] (WB, Human) WB Human
Sankowski R. Effects of the MAP kinase inhibitor CNI-1493 on Amyloid beta-treated primary and mikrogliare neuronal cells Thesis, Department of Medicine, Philipps-University Marburg. 2012 Nov (WB, Mouse) WB Mouse
Ramsingh G, Koboldt DC, Trissal M et al. Complete characterization of the microRNAome in a patient with acute myeloid leukemia. Blood. 2010 Dec 9 [PMID: 20876853] (WB, Human) WB Human

Reviews for GAPDH Antibody (NB300-285) (3) 4.73

Average Rating: 4.7
(Based on 3 reviews)
We have 3 reviews tested in 1 species: Human.

Reviews using NB300-285:
Filter by Applications
ICC
(2)
WB
(1)
All Applications
Filter by Species
Human
(3)
All Species
Images Ratings Applications Species Date Details
Immunocytochemistry GAPDH NB300-285
Enlarge
reviewed by:
Anony- mous
ICC Human 06/10/2014
View

Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsIF Confocal analysis of Hela cells using GAPDH antibody (NB615) (NB300-285, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF
Western Blot GAPDH NB300-285
Enlarge
reviewed by:
Anony- mous
WB Human 06/10/2014
View

Summary

ApplicationWestern Blot
SpeciesHuman
Special ApplicationsThis antibody detected endogenous levels of GAPDH proteins in Jurkat, HeLa and COS cell lysates in WB at dilution of 1:100.
FileView PDF
Immunocytochemistry GAPDH NB300-285
Enlarge
reviewed by:
Anony- mous
ICC Human 05/22/2014
View

Summary

ApplicationImmunocytochemistry
SpeciesHuman
FileView PDF

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 (NB300-285). (Showing 1 - 6 of 6 FAQs).

  1. We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
    • Well 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 lookin at. 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.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Allophycocyanin Labeled NB300-285APC
DyLight 350 Labeled NB300-285UV
DyLight 405 Labeled NB300-285V
DyLight 488 Labeled NB300-285G
DyLight 680 Labeled NB300-285FR
DyLight 755 Labeled NB300-285IR
PE Labeled NB300-285PE
PerCP Labeled NB300-285PCP

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Bioinformatics Tool for GAPDH Antibody (NB300-285)

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Diseases for GAPDH Antibody (NB300-285)

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Research Areas for GAPDH Antibody (NB300-285)

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Blogs on GAPDH. Showing 1-10 of 12 blog posts - Show all blog posts.

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 west blots. However, this pri...  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.

New Primers Available for ChIP Procedures
Novus has recently released six new primer sets designed for use in real-time PCR DNA amplification of housekeeping, silent, or heterochromatin associated proteins. Specifically, the primers are for GAPDH, RPL30, MyoD1, AFM, alpha Satellite, and Satel...  Read full blog post.

Showing 1-10 of 12 blog posts - Show all blog posts.

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Recent Reviews

4.7
5
2
4
1
3
0
2
0
1
0

Anony- mous
06/10/2014
Application: ICC
Species: Human

Anony- mous
06/10/2014
Application: WB
Species: Human

Anony- mous
05/22/2014
Application: ICC
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot