GAPDH Antibody (2D4A7) [HRP]

Images

 
Western Blot: GAPDH Antibody (2D4A7) [HRP] [NB300-328H] - Lysosome-associated membrane protein 2 (LAMP-2) upregulation with intracellular amyloid-B accumulations quantified against GAPDH. Image from verfied customer ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, RbSpecies Glossary
Applications WB, ELISA, IHC, IHC-P
Clone
2D4A7
Clonality
Monoclonal
Host
Mouse
Conjugate
HRP

GAPDH Antibody (2D4A7) [HRP] Summary

Immunogen
Ni-NTA purified recombinant human GAPDH expressed in E. Coli strain BL21 (DE3). [UniProt# P04406]
Marker
Cytosolic Marker
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
  • Western Blot
  • ELISA
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
Application Notes
This GAPDH antibody is useful for ELISA, Immunocytochemistry/Immunofluorescence, Immunohistochemistry paraffin embedded sections and Western Blot. Use in Immunoprecipitation and Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 25915564)

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.
Readout System
Reviewed Applications
Read 1 Review rated 5
using
NB300-328H in the following applications:

Publications
Read Publications using
NB300-328H in the following applications:

  • 1 publication
  • IP
    1 publication
  • WB
    9 publications

Reactivity Notes

Human, Mouse, Rabbit and Rat. Since GAPDH is one of the most conserved proteins known and it is therefore likely that the antibody will react with other species also. Porcine reactivity reported in scientific literature (PMID: 25063310)

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
PBS
Preservative
No Preservative
Purity
Protein G purified

Notes

This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for GAPDH Antibody (2D4A7) [HRP]

  • 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

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. It catalyzes an important energy-yielding step in carbohydrate metabolism, the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). The enzyme exists as a tetramer of identical chains. Besides its functioning as a glycolytic enzyme in cytoplasm, recent evidence suggest that mammalian GAPDH is also involved in a great number of intracellular proceses such as membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair. During the last decade a lot of findings appeared concerning the role of GAPDH in different pathologies including prostate cancer progression, programmed neuronal cell death, age- related neuronal diseases, such as Alzheimer's and Huntington's disease.

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.

Customers Who Viewed This Item Also Viewed...

NBP1-85789
Species: Hu
Applications: WB, ICC/IF, IHC, IHC-P
NB600-501
Species: Hu, Mu, Rt, Po, Bv, Ca, Ch, Fe, Fi, GP, Ha, Le, Ma, Pm, Rb, Sh, Sq, Ze
Applications: WB, Simple Western, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
NBP1-31470
Species: Hu, Mu, Rt
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-P
NB100-64792
Species: Hu, Mu, Rt, Rb
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr
NBP1-33527
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NB600-501
Species: Hu, Mu, Rt, Po, Bv, Ca, Ch, Fe, Fi, GP, Ha, Le, Ma, Pm, Rb, Sh, Sq, Ze
Applications: WB, Simple Western, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
NB100-236
Species: Hu, Mu
Applications: WB, IB, IHC, IHC-Fr, IHC-P, IP
NBP1-81838
Species: Hu, Mu
Applications: WB, IHC, IHC-P
H00026330-M01
Species: Hu
Applications: WB, ELISA, IHC, IHC-P
NB500-317
Species: Hu, Bv(-), Ca(-), Ch(-), Mu(-), Rb(-)
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-P, IP, RIA
NB300-605
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NBP2-24916
Species: Hu, Mu, Rt, Bv
Applications: WB, IHC, IHC-P
AF3589
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC
NBP2-34260
Species: Hu, Mu, Po, Bv, Rb
Applications: WB, Flow, ICC/IF, IHC-Fr, IHC-P, IP, PAGE
NBP2-25147
Species: Hu, Mu, Rt, Bv, Xp
Applications: WB, Simple Western, ICC/IF
NBP2-12933
Species: Hu, Mu, Rt, Po
Applications: WB, PEP-ELISA
NB600-41532
Species: Mu
Applications: WB, ELISA, ICC/IF, IHC, IHC-P

Publications for GAPDH Antibody (NB300-328H)(11)

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

We have publications tested in 3 applications: ICC/IF, IP, WB.


Filter By Application
ICC/IF
(1)
IP
(1)
WB
(9)
All Applications
Filter By Species
Human
(7)
Mouse
(1)
All Species
Showing Publications 1 - 10 of 11. Show All 11 Publications.
Publications using NB300-328H Applications Species
Bergstrom P, Agholme L, Nazir FH et al. Amyloid precursor protein expression and processing are differentially regulated during cortical neuron differentiation Sci Rep 2016 Jul 07 [PMID: 27383650] (WB) WB
Wei L, Wang H, Yang F et al. Interleukin-17 potently increases non-small cell lung cancer growth. Mol Med Rep 2016 Feb [PMID: 26708832]
Wang Y, Serricchio M, Jauregui M et al. Deubiquitinating enzymes regulate PARK2-mediated mitophagy Autophagy 2015 Apr 27 [PMID: 25915564] (WB, ICC/IF, IP, Human) WB, ICC/IF, IP Human
Carta S, Penco F, Lavieri R et al. Cell stress increases ATP release in NLRP3 inflammasome-mediated autoinflammatory diseases, resulting in cytokine imbalance Proc. Natl. Acad. Sci. U.S.A. 2015 Feb 17 [PMID: 25730877] (WB, Human) WB Human
Vene R, Cardinali B, Arena G et al. Glycogen Synthase Kinase 3 Regulates Cell Death and Survival Signaling in Tumor Cells under Redox Stress. Neoplasia. 2014 Sep 01 [PMID: 25246272]
Domert J, Rao SB, Agholme L et al. Spreading of amyloid-? peptides via neuritic cell-to-cell transfer is dependent on insufficient cellular clearance. Neurobiol. Dis. 2014 Jan 9 [PMID: 24412310] (WB, Human) WB Human
Amaro A, Mirisola V, Angelini G et al. Evidence of epidermal growth factor receptor expression in uveal melanoma: Inhibition of epidermal growth factor-mediated signalling by Gefitinib and Cetuximab triggered antibody-dependent cellular cytotoxicity. Eur J Cancer 2013 Jul 9 [PMID: 23849826] (WB, Human) WB Human
Carta S, Silvestri M, Rossi GA. Modulation of airway epithelial cell functions by Pidotimod: NF-kB cytoplasmatic expression and its nuclear translocation are associated with an increased TLR-2 expression. Ital J Pediatr. 2013 May 10 [PMID: 23663325] (WB, Human) WB Human
Benelli R, Stigliani S, Minghelli S et al. Impact of CXCL1 overexpression on growth and invasion of prostate cancer cell. Prostate 2013 Jan 17 [PMID: 23334998] (WB, Mouse) WB Mouse
Benelli R, Vene R, Minghelli S et al. Celecoxib induces proliferation and Amphiregulin production in colon subepithelial myofibroblasts, activating erk1-2 signaling in synergy with EGFR Cancer Lett 2012 Sep 22 [PMID: 23010081] (WB, Human) WB Human
Show All 11 Publications.

Review for GAPDH Antibody (NB300-328H) (1) 51

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

Reviews using NB300-328H:
Filter by Applications
WB
(1)
All Applications
Filter by Species
Human
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot GAPDH NB300-328H
Enlarge
reviewed by:
Sangeeta Nath
WB Human 05/08/2014
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate neuronal cells
SpeciesHuman
CommentsAntibody is extremely strong, therefore need high dilution (1:30,000) and very very short exposure to avoid over staining.

Blocking

Blocking Details5 % Dry milk in TBS-tween

Primary Anitbody

Dilution Ratio1:30,000 dilution in blocking buffer, incubation time 1 h at room temperature

Secondary Antibody

Secondary DescriptionRabbit anti-goat IgG, HRP
Secondary Concentration1:4000

Details

Detection Notes30 secs exposoure

Comments

CommentsAntibody is extremely strong, therefore need high dilution (1:30,000) and very very short exposure to avoid over staining.

Product General Protocols

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

Video Protocols

WB Video Protocol

FAQs for GAPDH Antibody (NB300-328H). (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.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Biotin Labeled NB300-328B
DyLight 350 Labeled NB300-328UV
DyLight 405 Labeled NB300-328V
DyLight 488 Labeled NB300-328G
DyLight 550 Labeled NB300-328R
DyLight 650 Labeled NB300-328C
DyLight 680 Labeled NB300-328FR
DyLight 755 Labeled NB300-328IR
FITC Labeled NB300-328F

Additional GAPDH Products

Bioinformatics Tool for GAPDH Antibody (NB300-328H)

Discover related pathways, diseases and genes to GAPDH Antibody (NB300-328H). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for GAPDH Antibody (NB300-328H)

Discover more about diseases related to GAPDH Antibody (NB300-328H).
 

Pathways for GAPDH Antibody (NB300-328H)

View related products by pathway.

PTMs for GAPDH Antibody (NB300-328H)

Learn more about PTMs related to GAPDH Antibody (NB300-328H).
 

Research Areas for GAPDH Antibody (NB300-328H)

Find related products by research area.

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.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Recent Reviews

5
5
1
4
0
3
0
2
0
1
0

Sangeeta Nath
05/08/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez