GAPDH Antibody

Images

 
Western Blot: GAPDH Antibody [NB300-326] - Detection of GAPDH in nu/nu Mouse cells. 5% BSA in PBS, 1:5,000. Secondary: Goat anti-Rabbit IgG 1:5,000. This image was submitted via customer Review.
Western Blot: GAPDH Antibody [NB300-326] - Detection of Mouse GAPDH Whole cell lysate (5, 15 and 50 ug) from mouse NIH3T3 cells. Affinity purified rabbit anti-GAPDH antibody used at 0.04 ug/ml
Simple Western: GAPDH Antibody [NB300-326] - 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 12-230kDa separation ...read more

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, Simple Western, IP (-)
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
0.2 mg/ml

Order Details

GAPDH Antibody Summary

Immunogen
The epitope recognized by this antibody maps to a region between residues 300 and the C-terminus (residue 335) of mouse GAPDH using the numbering given in entry NP_001001303.1 (GeneID 407972).
Marker
Cytosolic Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
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 1:5000
  • Simple Western 1:25
Application Notes
This GAPDH antibody is useful for Western blot, where a band is observed approx. 36 kDa. The antibody does not work in Immunoprecipitation.

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 1 Review rated 5
using
NB300-326 in the following applications:

Publications
Read Publications using
NB300-326 in the following applications:

  • WB
    1 publication

Reactivity Notes

Mouse. In tests of lysate from HeLa cells, reactivity with Human GAPDH was weak. Human reactivity reported in scientific literature (PMID: 27602114).

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
TBS and 0.1% BSA
Preservative
0.09% Sodium Azide
Concentration
0.2 mg/ml
Purity
Immunogen affinity purified

Alternate Names for GAPDH Antibody

  • 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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NB300-326
Species: Hu, Mu
Applications: WB, Simple Western, IP (-)

Publications for GAPDH Antibody (NB300-326)(2)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 1 application: WB.


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Review for GAPDH Antibody (NB300-326) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Mouse.

Reviews using NB300-326:
Filter by Applications
WB
(1)
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Filter by Species
Mouse
(1)
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Images Ratings Applications Species Date Details
Western Blot GAPDH NB300-326
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reviewed by:
Anonymous
WB Mouse 02/14/2017
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Summary

ApplicationWestern Blot
Sample TestedMOUSE FIBROBLAST
SpeciesMouse
LotA1

Product General Protocols

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WB Video Protocol

FAQs for GAPDH Antibody (NB300-326). (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

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Bioinformatics Tool for GAPDH Antibody (NB300-326)

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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

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1
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0
2
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1
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Anonymous
02/14/2017
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot