GAPDH Antibody (1A10)

Images

 
Western Blot: GAPDH Antibody (1A10) [NBP1-47339] - Western blot analysis using anti-GAPDH mAb against Hela (1), A549 (2), A431 (3), MCF-7 (4), K562 (5), Jurkat (6), HL60 (7), SKN-SH (8) and SKBR-3 (9) cell lysate.
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody (1A10) [NBP1-47339] - GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using 2 ug/mL of mouse anti-GAPDH monoclonal ...read more
Immunohistochemistry-Paraffin: GAPDH Antibody (1A10) [NBP1-47339] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma (left) and kidney carcinoma (right), showing cytoplasmic localization using ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody (1A10) [NBP1-47339] - Confocal immunofluorescence analysis of methanol-fixed HepG2 (left) and Hela (right) cells using anti-GAPDH mAb (green), showing cytoplasmic ...read more
Simple Western: GAPDH Antibody (1A10) [NBP1-47339] - 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, MuSpecies Glossary
Applications WB, Simple Western, ELISA, ICC/IF, IHC, IHC-P
Clone
1A10
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml

GAPDH Antibody (1A10) Summary

Immunogen
Purified recombinant fragment of human GAPDH expressed in E. coli. [UniProt# P04406]
Localization
Cytoplasmic
Marker
Cytosolic Marker
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
Purity
Ammonium sulfate precipitation
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 1:500-1:2000
  • Simple Western 1:50
  • ELISA 1:10000
  • Immunocytochemistry/Immunofluorescence 1:200-1:1000
  • Immunohistochemistry 1:200-1:1000
  • Immunohistochemistry-Paraffin 1:200-1:1000
Application Notes
This GAPDH (1A10) antibody is useful for Western blot, Immunohistochemistry on paraffin embedded sections, Immunocytochemistry/Immunofluorescence and ELISA.

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.
Positive Control
Jurkat Lysate (NB800-PC2)
A-431 Lysate (NB800-PC12)
HeLa Lysate (NB800-PC1)
MCF-7 Lysate (NB820-59465)
Reviewed Applications
Read 2 Reviews rated 4.5
using
NBP1-47339 in the following applications:

Publications
Read Publications using
NBP1-47339 in the following applications:

  • WB
    3 publications

Reactivity Notes

Human. Mouse reactivity reported in scientific literature (PMID: 25833141)

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.03% Sodium Azide
Concentration
1.0 mg/ml
Purity
Ammonium sulfate precipitation

Alternate Names for GAPDH Antibody (1A10)

  • 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 is a 146 kDa tetramer composed of four 30-40 kDa subunits. Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a metabolic enzyme responsible for catalyzing one step in the glycolytic pathway, the reversible oxidative phosphorylation of glyceraldehyde 3-phosphate. Because GAPDH as a protein expressed in large amounts and which is required at all times for an important house keeping functions, levels of GAPDH mRNA are often used as standards in studies of mRNA expression. Increasingly, scientists are making use of specific antibodies to GAPDH as loading controls for western blotting experiments. Apart from a role in glycolysis, GAPDH may have other roles such as in the activation of transcription. GAPDH is reported to bind to a variety of other proteins, including the amyloid precursor protein, mutations in which cause some forms of Alzheimer's disease, and the polyglutamine tracts of Huntingtin, the protein product aberrant forms of which are causative of Huntington's disease. Associations with actin and tubulin have also be reported. The protein may also have a role in the regulation of apoptosis, and interestingly migrates from the cytoplasm into the nucleus when cells become apoptotic.

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 (NBP1-47339)(4)

Reviews for GAPDH Antibody (NBP1-47339) (2) 4.52

Average Rating: 4.5
(Based on 2 reviews)
We have 2 reviews tested in 1 species: Human.
We have 2 reviews tested in 1 application: WB.

Reviews using NBP1-47339:
Filter by Applications
WB
(2)
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Filter by Species
Human
(2)
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Images Ratings Applications Species Date Details
 
reviewed by:
Anonymous
WB Human 12/23/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 24421388
SpeciesHuman

Blocking

Blocking DetailsSee PMID 24421388

Details

Detection NotesSee PMID 24421388

Comments

CommentsIndependent review from antibodyresource.comPublished in PMID 24421388.
Western Blot GAPDH NBP1-47339
Enlarge
reviewed by:
Ranveer Singh
WB Human 06/02/2014
View

Summary

ApplicationWestern Blot
Sample TestedHeLa cells
SpeciesHuman
CommentsA good antibody. Can be reused multiple times.

Blocking

Blocking Details5% BSA in TBST for 1 h

Primary Anitbody

Dilution Ratio1:3000, incubated 3h at RT in 5% BSA in TBST

Secondary Antibody

Secondary DescriptionGoat anti-mouse IgG HRP (Biorad)
Secondary Manufacturer Cat#170-5047
Secondary Concentration1:10,000

Details

Detection NotesECL reagent was used. Exposure for 10 seconds

Comments

CommentsA good antibody. Can be reused multiple times.

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 (NBP1-47339). (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.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

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Bioinformatics Tool for GAPDH Antibody (NBP1-47339)

Discover related pathways, diseases and genes to GAPDH Antibody (NBP1-47339). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
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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

4.5
5
1
4
1
3
0
2
0
1
0

 
Anonymous
12/23/2014
Application: WB
Species: Human

Ranveer Singh
06/02/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot