GAPDH Antibody (13H12)

Images

 
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.
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
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
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - WB detection of GAPDH protein in HeLa cells lysate using GAPDH antibody (clone 13H12) in (A) the absence and (B) the presence of immunizing peptide.
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - WB detection of GAPDH protein in lysates of Mouse cell lines (A) NIH 3T3 (B) RAW 264.7 using GAPDH antibody (clone 13H12) at a concentration of 0.25 ug/ml.
Western Blot: GAPDH Antibody (13H12) [NBP2-27103] - WB detection of GAPDH protein in lysates of (A) HEK 293 cells and (B) Drosophila larvae using GAPDH antibody (clone 13H12) at a concentration of 0.25 ug/ml.
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
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/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 under reducing conditions using the 12-230 ...read more

Product Details

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

GAPDH Antibody (13H12) Summary

Immunogen
Amino acids between 275 and 325 of glyceraldehyde 3-phosphate dehydrogenase protein were used as the immunogen for this antibody.
Localization
Cytoplasm, Nucleus
Marker
Cytosolic Marker
Predicted Species
Porcine (91%), Bovine (91%), Guinea Pig (91%), Ferret (91%), Reptile (91%), Canine (91%), Squirrel (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
  • Western Blot 0.25 - 1 ug/ml
  • Simple Western 1:25
  • Immunocytochemistry/Immunofluorescence 1:10
  • Immunohistochemistry 5 ug/ml
  • Immunohistochemistry-Paraffin 5 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.
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
GAPDH Lysate (H00002597-T01)
GAPDH Lysate (NBL1-10967)
Various Tissue Micro Array (NBP2-30234)
Reviewed Applications
Read 3 Reviews rated 5
using
NBP2-27103 in the following applications:

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

  • WB
    18 publications

Reactivity Notes

Reacts with Human, Primates/Monkey, Mouse and Drosophila. 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 reactivity reported in scientific literature (PMID: 24796753). Sheep reactivity reported in scientific literature (PMID: 27618403).

Packaging, Storage & Formulations

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

Alternate Names for GAPDH Antibody (13H12)

  • 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) catalyzes the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD), an important energy-yielding step in carbohydrate metabolism. Recent evidence suggests that it also is involved in a number of cellular processes such as membrane fusion, phosphotransferase activity, DNA replication and repair, and nuclear RNA export. GAPDH has also been implicated in playing a role in different pathologies such as cancer progression, apoptosis, and neuronal diseases such as Alzheimer's and Huntington's disease. GAPDH is constitutively expressed at high levels in almost all tissues and cell lines making it ideal for use as a loading control marker in immunoblots.

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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Supplier Logo

Publications for GAPDH Antibody (NBP2-27103)(19)

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

We have publications tested in 1 application: WB.


Filter By Application
WB
(18)
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Filter By Species
Human
(8)
Mouse
(5)
Rat
(2)
Drosophila
(1)
Monkey
(1)
Primate
(2)
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(1)
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Showing Publications 1 - 10 of 19. Show All 19 Publications.
Publications using NBP2-27103 Applications Species
Krassovka JM. inaugural-Dissertation zur Erlangung des Doktogrades Thesis 2017 (WB) WB
Winzi M, Casas Vila N, Paszkowski-Rogacz M, Ding L. The long noncoding RNA lncR492 inhibits neural differentiation of murine embryonic stem cells. PLoS ONE. 2018 Jan 24 [PMID: 29364956] (WB) WB
Yeyati PL, Schiller R, Mali G et al. KDM3A coordinates actin dynamics with intraflagellar transport to regulate cilia stability. J. Cell Biol. Feb 28 2017 [PMID: 28246120] (WB, Mouse) WB Mouse
Bologna-Molina R, Takeda Y, Kuga T et al. Expression of Wilms' tumor 1 (WT1) in ameloblastomas J Oral Sci Sep 26 2016 [PMID: 27665981] (WB, Human) WB Human
Titos E, Rius B, Lopez-Vicario C et al. Signaling and Immunoresolving Actions of Resolvin D1 in Inflamed Human Visceral Adipose Tissue J. Immunol. Sep 19 2016 [PMID: 27647830] (WB, Human) WB Human
Mishra D, Singh S, Narayan G, Curcumin Induces Apoptosis in Pre-B Acute Lymphoblastic Leukemia Cell Lines Via PARP-1 Cleavage Asian Pac. J. Cancer Prev. Sep 20 2016 [PMID: 27644631] (WB, Human) WB Human
Spasova MS, Chen X, Sadowska GB et al. Ischemia Reduces Inter-Alpha Inhibitor Proteins in the Brain of the Ovine Fetus Dev Neurobiol Sep 12 2016 [PMID: 27618403] (WB, Sheep) WB Sheep
Grimberg-Peters D, Buren C, Windolf J et al. Hyperbaric Oxygen Reduces Production of Reactive Oxygen Species in Neutrophils from Polytraumatized Patients Yielding in the Inhibition of p38 MAP Kinase and Downstream Pathways PLoS ONE Aug 17 2016 [PMID: 27529549] (WB, Human) WB Human
Srinivas C, Ramaiah MJ, Lavanya A et al. Novel Etoposide Analogue Modulates Expression of Angiogenesis Associated microRNAs and Regulates Cell Proliferation by Targeting STAT3 in Breast Cancer. PLoS ONE. 2015 Nov 10 [PMID: 26551008] (WB, Human) WB Human
Sun S, Reddy BV, Irvine KD. Localization of Hippo signalling complexes and Warts activation in vivo. Nat Commun. 2015 Sep 30 [PMID: 26420589] (WB, Drosophila)

Details:
GAPDH antibody clone 13H12 was used as WB loading control in experiments involving S2 cells (Schneider 2 cells) which were subjected to transfection using plasmids pAC5.1-Wts:V5 (D. Pan), pAC5.1-HA:Ex (G. Halder), pAC5.1-Myc:Hpo (D. Pan), pUAST-Flag:Mats, aw-Gal4 or pAC5.1-Flag:GFP28.
WB Drosophila
Show All 19 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)
All Applications
Filter by Species
Human
(1)
Mouse
(1)
Other
(1)
All Species
Images Ratings Applications Species Date Details
 
reviewed by:
Michaela Rekow
WB Mouse 01/14/2016
View

Summary

ApplicationWestern Blot
Sample TestedSplenocytes
SpeciesMouse

Blocking

Blocking Details5% Milk, 45 Minutes, Room Temp

Primary Anitbody

Dilution Ratio1:10,000

Secondary Antibody

Secondary DescriptionAnti-rabbit HRP

Details

Detection NotesDeveloper, Exposure times from 30 seconds - 15 minutes
 
reviewed by:
Anonymous
WB Other 12/29/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 24465661
SpeciesOther

Blocking

Blocking DetailsSee PMID 24465661

Details

Detection NotesSee PMID 24465661

Comments

CommentsPublished in PMID 24465661.Independent review from antibodyresource.com
Western Blot GAPDH NBP2-27103
Enlarge
reviewed by:
Anonymous
WB Human 10/09/2014
View

Summary

ApplicationWestern Blot
Sample TestedHeLa and HEK 293 cells
SpeciesHuman

Blocking

Blocking Details4 % BSA in TBST

Primary Anitbody

Dilution RatioDiluted 1:2000 times

Secondary Antibody

Secondary DescriptionGoat Anti-Mouse IgG HRP Affinity Purified PAb (R&D Systems)
Secondary Manufacturer Cat#HAF007
Secondary ConcentrationDiluted 5,000 times

Details

Detection NotesOne-hour Western detection system (Genscript), 10 sec exposure

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 - 7 of 7 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.
  7. 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.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NBP2-27103AF405
Alexa Fluor 488 Labeled NBP2-27103AF488
Alexa Fluor 647 Labeled NBP2-27103AF647
Alexa Fluor 700 Labeled NBP2-27103AF700
Biotin Labeled NBP2-27103B
DyLight 350 Labeled NBP2-27103UV
DyLight 405 Labeled NBP2-27103V
DyLight 488 Labeled NBP2-27103G
DyLight 550 Labeled NBP2-27103R
DyLight 650 Labeled NBP2-27103C
DyLight 680 Labeled NBP2-27103FR
DyLight 755 Labeled NBP2-27103IR
HRP Labeled NBP2-27103H

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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

5
5
3
4
0
3
0
2
0
1
0

 
Michaela Rekow
01/14/2016
Application: WB
Species: Mouse

 
Anonymous
12/29/2014
Application: WB
Species: Other

Anonymous
10/09/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot