We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?

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 interested in. GAPDH antibody is a good choice for low to mid MW targets, as GAPDH detects around 35 kDa.

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.

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.

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.

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.

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.

I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.

For homogenized tissue, beta-actin and GAPDH are both fine.

We received GAPDH antibody NB600-502 and there was only about 15µl of antibody in the tube. I was under the impression this should have been 100µl.

The unit size of this product is 0.1 mg with a concentration of 6.7 mg/ml. Hence the volume should be no less than 14.9 ul.

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GAPDH Antibody (2D4A7)

Name: GAPDH Antibody (2D4A7)
Brand: Novus Biologicals
SKU: NB300-328
Rating: 5 (12 reviews)

(68 Publications)

(12 Reviews)

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Product Details

Summary

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Catalog Number

NB300-328
Availability

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GAPDH Antibody (2D4A7) Summary

Immunogen	GAPDH antibody raised against 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	Ammonium sulfate precipitation
Innovator's Reward	Test in a species/application not listed above to receive a full credit towards a future purchase. Learn about the Innovator's Reward

Applications/Dilutions

Dilutions

ELISA 1:100-1:2000
Immunocytochemistry/ Immunofluorescence 1:200-1:1000
Immunohistochemistry 1:1000-1:4000
Immunohistochemistry-Paraffin 1:1000-1:4000
Immunoprecipitation
Simple Western 1:250
Western Blot 1:20000

Application Notes

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. Use in immunoprecipitation reported in scientific literature (PMID: 25915564).

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 Overexpression Lysate

Reviewed Applications

Read 12 Reviews rated 5

using
NB300-328 in the following applications:

Western Blot
Immunocytochemistry

Publications

Read Publications using
NB300-328 in the following applications:

FLOW
1 publication
ICC/IF
2 publications
IP
1 publication
WB
57 publications
Western Blot
1 publication

Reactivity Notes

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

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 (2D4A7)

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

Background

Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis, converting glyceraldehyde-3-phosphate into 1,3 diphosphoglycerate. This constitutively expressed, homotetramer protein can be found in the nucleus and cytoplasm, and the monomer has a theoretical molecular weight of 36 kDa. Known as a housekeeping gene, GAPDH routinely serves as a control for real-time PCR (RT-PCR) and as a loading control for Western Blots (1-3). In addition to its role in glycolysis, GAPDH has multiple cellular functions including membrane trafficking, transcription activation, apoptosis, DNA repair, and has been implicated in various pathologies such as cancer and neurodegenerative diseases including Alzheimer's and Huntington's (4,5).

References

1) Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3):389-95. PMID: 15769908

2) Jia Y, Takimoto K. (2006) Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res. 98(3):386-93. PMID: 16385079

3) Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew TL, Green KJ. (2005) Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol. 171(6):1045-59. PMID: 16365169

4) Sirover MA1. (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1432(2): 159-84. PMID: 10407139

5) Tristan C, Shahani N, Sedlak TW, Sawa A. (2011) The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal. 23(2):317-23. PMID: 20727968

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 (NB300-328)(68)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Porcine, Rabbit.

We have publications tested in 5 applications: FLOW, ICC/IF, IP, WB, Western Blot.

Showing Publications 1 - 10 of 68. Show All 68 Publications.

Publications using NB300-328	Applications	Species
Eriksson I, Vainikka L, W�ster P, �llinger K Lysosomal function and intracellular position determine the malignant phenotype in malignant melanoma The Journal of investigative dermatology 2023-03-03 [PMID: 36871659] (Western Blot, Human)	Western Blot	Human
Mukherjee A, Islam S, Kieser RE et al. Long noncoding RNAs are substrates for cytoplasmic capping enzyme FEBS letters 2023-03-01 [PMID: 36856012]
Demers ND, Riccio V, Jo DS et al. PEX13 prevents pexophagy by regulating ubiquitinated PEX5 and peroxisomal ROS Autophagy 2023-06-01 [PMID: 36541703] (WB, Human)	WB	Human
Sharma V, Thakore P, Majumdar S THAP9 Transposase Cleaves DNA via Conserved Acidic Residues in an RNaseH-Like Domain Cells 2021-05-29 [PMID: 34072453] (WB)	WB
Camporesi E, Lashley T, Gobom J et al. Neuroligin-1 in brain and CSF of neurodegenerative disorders: investigation for synaptic biomarkers Acta neuropathologica communications 2021-02-01 [PMID: 33522967] (WB, Human)	WB	Human
Unsicker C, Cristian FB, von Hahn M, et al. SHANK2 mutations impair apoptosis, proliferation and neurite outgrowth during early neuronal differentiation in SH-SY5Y cells Scientific reports 2021-01-22 [PMID: 33483523] (WB, Human)	WB	Human
Hung WT, Wang CH, Lin SY et al. Leptin protects brain from ischemia/reperfusion-induced infarction by stabilizing the blood-brain barrier to block brain infiltration of the blood-borne neutrophils Eur. J. Neurosci. 2020-07-07 [PMID: 32638449]
Eriksson I, WAster P, Ollinger K Restoration of lysosomal function after damage is accompanied by recycling of lysosomal membrane proteins Cell Death Dis 2020-05-14 [PMID: 32409651] (WB, Human)	WB	Human
Lachenauer, E R, Stabler, S P Et al. p53 Disruption Increases Uracil Accumulation in DNA of Murine Embryonic Fibroblasts and Leads to Folic Acid-Nonresponsive Neural Tube Defects in Mice. J Nutr 2020-07-01 [PMID: 32271909] (FLOW, Human)	FLOW	Human
Wang S, Lin M, Hsu F et al. Synthesis and Biological Evaluation of 2-Quinolineacrylamides Bioorganic & Medicinal Chemistry 2019-12-01 [PMID: 31924504] (WB, Human)	WB	Human
Cvitic S, Strutz J, Appel HM Sexual dimorphism of miRNA signatures in feto-placental endothelial cells is associated with altered barrier function and actin organisation Clin. Sci. 2019-12-11 [PMID: 31825070]
Shea J, Terry C, Edwards K, Agarwal J Glitazone loaded fat enhances adiponectin production and inhibits breast cancer cell proliferation Mol. Biol. Rep. 2019-09-30 [PMID: 31571108] (WB, Human)	WB	Human
Zhang Q, Narayanan V, Mui KL et al. Mechanical Stabilization of the Glandular Acinus by Linker of Nucleoskeleton and Cytoskeleton Complex Curr. Biol. 2019-08-02 [PMID: 31402305]
Wu TT, Chen CC, Lin JT et al. The anti-inflammatory function of adenine occurs through AMPK activation and its downstream transcriptional regulation in THP-1 cells Biosci. Biotechnol. Biochem. 2019-08-08 [PMID: 31392929] (ICC/IF)	ICC/IF
McSweeney KM, Bozza WP, Alterovitz WL et al. Transcriptomic profiling reveals p53 as a key regulator of doxorubicin-induced cardiotoxicity Cell Death Discov 2019-06-12 [PMID: 31231550] (WB, Human)	WB	Human
Shen CL, Huang WH, Hsu HJ et al. GAP31 from an ancient medicinal plant exhibits anti-viral activity through targeting to Epstein-Barr virus nuclear antigen 1 Antiviral Res. 2019-02-25 [PMID: 30817940] (WB, Human)	WB	Human
Huguier V, Giot JP, Simonneau M et al. Oncostatin M exerts a protective effect against excessive scarring by counteracting the inductive effect of TGFb1 on fibrosis markers Sci Rep 2019-02-14 [PMID: 30765798] (WB, Human)	WB	Human
Riccio V, Demers N, Hua R et al. Deubiquitinating enzyme USP30 maintains basal peroxisome abundance by regulating pexophagy J. Cell Biol. [PMID: 30700497] (WB, Human)	WB	Human
Melissaridou S, Wiechec E, Magan M et al. The effect of 2D and 3D cell cultures on treatment response, EMT profile and stem cell features in head and neck cancer Cancer Cell Int Cancer Cell Int. 2019-01-14 [PMID: 30651721] (WB, Human) Details: Citation using the HRP form of this antibody.	WB	Human
Twomey JD, Zhao L, Luo S et al. Tubulin couples death receptor 5 to regulate apoptosis. Oncotarget 2018-12-04 [PMID: 30613368] (WB, Human)	WB	Human
Sexton RE, Hachem AH, Assi AA et al. Metabotropic glutamate receptor-1 regulates inflammation in triple negative breast cancer. Sci Rep. 2018-10-30 [PMID: 30375476] (WB, Human)	WB	Human
Ehyai S, Miyake T, Williams D et al. FMRP recruitment of B-catenin to the translation pre-initiation complex represses translation. EMBO Rep. 2018-10-25 [PMID: 30361391] (WB, Human)	WB	Human
Carrasco-Ruiz MLa, Hernandez-Aragon LG, Chavez-Rios JR et al. High Estradiol Differentially Affects the Expression of the Glucose Transporter Type 4 in Pelvic Floor Muscles of Rats. Int Neurourol J. 2018-09-01 [PMID: 30286578] (WB, Rat)	WB	Rat
Cao J, Qiu J, Wang X et al. Identification of microRNA-124 in regulation of Hepatocellular carcinoma through BIRC3 and the NF-kB pathway J. Cancer 2018-08-12 [PMID: 30210622] (WB, Human)	WB	Human
Bozza WP, Zhang Y, Zhang B. Cytokeratin 8/18 protects breast cancer cell lines from TRAIL-induced apoptosis Oncotarget 2018-05-01 [PMID: 29796187] (WB, Human)	WB	Human
Prasad CB, Prasad SB, Yadav SS et al. Olaparib modulates DNA repair efficiency, sensitizes cervical cancer cells to cisplatin and exhibits anti-metastatic property Sci Rep. 2017-10-08 [PMID: 28993682] (WB, Human)	WB	Human
Cao J, Yan XR, Liu T et al. MicroRNA-552 promotes tumor cell proliferation and migration by directly targeting DACH1 via the Wnt/B-catenin signaling pathway in colorectal cancer Oncol Lett 2017-09-01 [PMID: 28927149] (WB, Human)	WB	Human
Speyer CL, Bukhsh MA, Jafry WS et al. Riluzole synergizes with paclitaxel to inhibit cell growth and induce apoptosis in triple-negative breast cancer Breast Cancer Res. Treat. 2017-08-05 [PMID: 28780701] (WB, Human)	WB	Human
Waster P, Orfanidis K, Eriksson I et al. UV radiation promotes melanoma dissemination mediated by the sequential reaction axis of cathepsins-TGF-b1-FAP-a Br. J. Cancer 2017-08-08 [PMID: 28697174] (WB, Human)	WB	Human
Law KB, Bronte-Tinkew D, Di Pietro E et al. The peroxisomal AAA ATPase complex prevents pexophagy and development of peroxisome biogenesis disorders Autophagy. 2017-05-04 [PMID: 28521612] (WB, Human) Details: Citation used the HRP format of this antibody.	WB	Human
Zocchi MR, Costa D, Vene R et al. Zoledronate can induce colorectal cancer microenvironment expressing BTN3A1 to stimulate effector gammadelta T cells with antitumor activity. Oncoimmunology. 2017-01-06 [PMID: 28405500] (WB, Human) Details: This citation used the HRP form of this antbody	WB	Human
Zhao L, Zhang B. Doxorubicin induces cardiotoxicity through upregulation of death receptors mediated apoptosis in cardiomyocytes. Sci Rep 2017-03-16 [PMID: 28300219] (WB, Human)	WB	Human
Bulanova Daria R, Akimov Yevhen A, Rokka Anne et al. Orphan G protein-coupled receptor GPRC5A modulates integrin beta 1-mediated epithelial cell adhesion. Cell Adhesion & Migration 2017-02-06 [PMID: 27715394] (WB, Human)	WB	Human
Bergstrom P, Agholme L, Nazir FH et al. Amyloid precursor protein expression and processing are differentially regulated during cortical neuron differentiation. Sci Rep. 2016-07-07 [PMID: 27383650] (WB) Details: This citation used the HRP version of this antibody.	WB
Lee HY, Chang CY, Su CJ et al. 2-(Phenylsulfonyl)quinoline N-hydroxyacrylamides as potent anticancer agents inhibiting histone deacetylase. Eur J Med Chem 2016-06-16 [PMID: 27344487] (WB, Human)	WB	Human
Speyer CL, Nassar MA, Hachem AH et al. Riluzole mediates anti-tumor properties in breast cancer cells independent of metabotropic glutamate receptor-1. Breast Cancer Res. Treat. 2016-06-01 [PMID: 27146584] (WB, Human)	WB	Human
Wei L, Wang H, Yang F et al. Interleukin-17 potently increases non-small cell lung cancer growth. Mol Med Rep. 2016-02-01 [PMID: 26708832] Details: This citation used the HRP version of this antibody.
Wang Y, Serricchio M, Jauregui M et al. Deubiquitinating enzymes regulate PARK2-mediated mitophagy. Autophagy 2015-04-27 [PMID: 25915564] (IP, WB, ICC/IF, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	IP, WB, ICC/IF	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 USA 2015-02-17 [PMID: 25730877] (WB, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	WB	Human
Cuellar TL, Barnes D, Nelson C et al. Systematic evaluation of antibody-mediated siRNA delivery using an industrial platform of THIOMAB-siRNA conjugates. Nucleic Acids Res. 2014-12-30 [PMID: 25550431] (WB, Human)	WB	Human
Field MS, Kamynina E, Watkins D et al. Human mutations in methylenetetrahydrofolate dehydrogenase 1 impair nuclear de novo thymidylate biosynthesis. Proc. Natl. Acad. Sci. U.S.A. 2014-12-29 [PMID: 25548164] (WB, Human)	WB	Human
Salcher S, Hagenbuchner J, Geiger K et al. C10ORF10/DEPP, a transcriptional target of FOXO3, regulates ROS-sensitivity in human neuroblastoma. Mol. Cancer. 2014-10-14 [PMID: 25261981] (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. 2014-07-31 [PMID: 25246272]
Lin YS, Yang CC, Hsu CC et al. Establishment of a novel, eco-friendly transgenic pig model using porcine pancreatic amylase promoter-driven fungal cellulase transgenes. Transgenic Res. 2014-07-26 [PMID: 25063310] (WB, Porcine) Details: GAPDH antibody used at 1:20,000 dilution in WB assay for detection of housekeeping protein GAPDH /loading control on pancreas of transgenic pig/fungal cellulase transgene expression driven by the porcine pancreatic amylase promoter in pigs and nontransgenic littermate (Fig 4 B).	WB	Porcine
Poenitzsch AM, Setaluri V, Spiegelman VS. microRNA-340 as a modulator of RAS-RAF-MAPK signaling in melanoma. Arch. Biochem. Biophys. 2014-07-17 [PMID: 25043973] (WB, Human) Details: GAPDH antibody used for WB on lysates of melanoma cells 451 Lu transfected or treated with - Lane 1 miRZip scrambled hairpin RNA control, Lane 2 miR-340 inhibiting miRZip340, Lane 3 PCMV-MIR over expression control, Lane 4 PCMV-MIR340 miR-340 over expression, Lane 5 DMSO control, Lane 6 50 mM MEK1 inhibitor PD98059 (Fig. 4a). Antibody also used on Hs294T, 451 Lu and SK-MEL-2 cells transfected with miRZip Scrambled or miRZip340 (Supplemental Figure 1).	WB	Human
Speyer CL, Hachem AH, Assi AA et al. Metabotropic glutamate receptor-1 as a novel target for the antiangiogenic treatment of breast cancer. PLoS ONE 2014-03-17 [PMID: 24633367] (WB, Human)	WB	Human
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 2014-01-09 [PMID: 24412310] (WB, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	WB	Human
Banda M, Speyer CL, Semma SN et al. Metabotropic glutamate receptor-1 contributes to progression in triple negative breast cancer. PLoS ONE 2014-01-09 [PMID: 24404125] (WB, Human)	WB	Human
Borghgraef P, Menuet C, Theunis C et al. Increasing Brain Protein O-GlcNAc-ylation Mitigates Breathing Defects and Mortality of Tau.P301L Mice. PLoS One 2013-12-23 [PMID: 24376810] (WB, Mouse)	WB	Mouse
Lin HP, Lin CY, Hsiao PH et al. Difference in Protein Expression Profile and Chemotherapy Drugs Response of Different Progression Stages of LNCaP Sublines and Other Human Prostate Cancer Cells. PLoS One 2013-12-05 [PMID: 24349321] (WB, Human)	WB	Human
Lin H, Hsu FC, Baumann BH et al. Cortical synaptic NMDA receptor deficits in A7 nicotinic acetylcholine receptor gene deletion models: Implications for neuropsychiatric diseases. Neurobiol Dis 2014-03-01 [PMID: 24326163] (WB, Mouse)	WB	Mouse
Wang YI, Bettaieb A, Sun C et al. Triglyceride-rich lipoprotein modulates endothelial vascular cell adhesion molecule (VCAM)-1 expression via differential regulation of endoplasmic reticulum stress. PLoS One. [PMID: 24205197] (WB, Human)	WB	Human
Deverse JS, Sandhu AS, Mendoza N et al. Shear stress modulates VCAM-1 expression in response to TNF-alpha and dietary lipids via interferon regulatory factor-1 in cultured endothelium. Am J Physiol Heart Circ Physiol. 2013-10-01 [PMID: 23934855] (WB, Porcine)	WB	Porcine
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 2013-07-09 [PMID: 23849826] (WB, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	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 2013-05-10 [PMID: 23663325] (WB, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	WB	Human
Shin M, Mizokami A, Kim J et al. Exogenous SPARC suppresses proliferation and migration of prostate cancer by interacting with integrin beta1 Prostate 2013-03-26 [PMID: 23532895] (WB, Human)	WB	Human
Peng HY, Chen GD, Lai CY et al. Spinal Serum-Inducible and Glucocorticoid-Inducible Kinase 1 Mediates Neuropathic Pain via Kalirin and Downstream PSD-95-Dependent NR2B Phosphorylation in Rats J Neurosci 2013-03-20 [PMID: 23516288] (WB, Rat)	WB	Rat
Storm P, Klausen TK, Trulsson M et al. A unifying mechanism for cancer cell death through ion channel activation by HAMLET. PLoS One. 2013-01-01 [PMID: 23505537] (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 2013-01-17 [PMID: 23334998] (WB, Mouse) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	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 2012-09-22 [PMID: 23010081] (WB, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	WB	Human
Loke WM, Lam KM, Chong WL et al. Products of 5-lipoxygenase and myeloperoxidase activities are increased in young male cigarette smokers Free Radic Res 2012-07-04 [PMID: 22690830] (WB, Human)	WB	Human
Deverse JS, Bailey KA, Jackson KN, Passerini AG. Shear stress modulates RAGE-mediated inflammation in a model of diabetes-induced metabolic stress Am J Physiol Heart Circ Physiol 2012-06-01 [PMID: 22467309] (WB, Human)	WB	Human
Field MS, Anderson DD, Stover P et al. Mthfs is an Essential Gene in Mice and a Component of the Purinosome Front Genet 2011-01-01 [PMID: 22303332] (WB, Mouse)	WB	Mouse
Andersson M, Lutay N, Hallgren O, Westergren-Thorsson G, Svensson M, Godaly G. Mycobacterium bovis bacilli Calmette-Guerin regulates leukocyte recruitment by modulating alveolar inflammatory responses. Innate Immun. 2012-01-12 [PMID: 22058091] (WB, Human)	WB	Human
Ciarlo M, Benelli R, Barbieri O, Minghelli S, Barboro P, Balbi C, Ferrari N. Regulation of neuroendocrine differentiation by AKT/hnRNPK/AR/Beta-catenin signaling in prostate cancer cells. Int J Cancer 2012-08-01 [PMID: 22015967] (WB, Human) Details: Using the HRP conjugated version of NB300-328, catalog number NB300-328H.	WB	Human
Gao J, Cai Q, Lu J et al. Upregulation of Cellular Bcl-2 by the KSHV Encoded RTA Promotes Virion Production. PLoS ONE 20116(8). [PMID: 21901143]
Johnen S, Kazanskaya O, Armogan N et al. Endogenic regulation of proliferation and zinc transporters by pigment epithelial cells nonvirally transfected with PEDF Invest Ophthalmol Vis Sci 2011-07-23 [PMID: 21508100] (WB, Rabbit)	WB	Rabbit
Cheng, G et al. Involvement of chloride channels in TGF-beta1-induced apoptosis of human bronchial epithelial cells. Am J Physiol Lung Cell Mol Physiol293(5):L1339-47. 2007-11-01 [PMID: 17873007]
Show All 68 Publications. Collapse Publications.

Reviews for GAPDH Antibody (NB300-328) (12) 512

Average Rating: 5

(Based on 12 reviews)

We have 12 reviews tested in 4 species: Human, Mouse, Human and Mouse, Pig.

Reviews using NB300-328:

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(9)

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(3)

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Human

(9)

Mouse

(1)

Human and Mouse

(1)

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reviewed by:
Verified Customer

Human

04/04/2020

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Summary

Application	Western Blot
Sample Tested	A2780 ovarian cancer cells,hela cell
Species	Human
Lot	E-3

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reviewed by:
Verified Customer

Human

12/27/2019

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Summary

Application	Western Blot
Sample Tested	A549 cells
Species	Human
Lot	E-3

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reviewed by:
Verified Customer

Human and Mouse

02/19/2019

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Summary

Application	Western Blot
Sample Tested	B16-F1 mouse melanoma cell line,HCC827 human non-small cell lung cancer cell line
Species	Human and Mouse
Lot	E-3

reviewed by:
Verified Customer

Human

01/31/2019

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Summary

Application	Western Blot
Sample Tested	Breast cancer cells
Species	Human
Lot	E-1

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reviewed by:
Verified Customer

Mouse

10/28/2018

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Application	Western Blot
Sample Tested	ct26,EMT6 cells
Species	Mouse
Lot	E-3

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reviewed by:
Verified Customer

Human

07/17/2018

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Summary

Application	Western Blot
Sample Tested	Breast cancer cells
Species	Human
Lot	E-1

Comments

Total cell lysates from T47D, MDA-MB-231, MCF-7 and MCF-10A cells were subjected to western blot. PVDF membrane were probed with 0.2 mm/ml Human GAPDH Antibody (2D4A7) (NB300-328). A specific band was detected for GAPDH at approximately 37 kDa. This experiment was conducted under reducing conditions

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reviewed by:
Verified Customer

ICC

Pig

01/04/2017

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Summary

Application	Immunocytochemistry
Sample Tested	Pig TM cells
Species	Pig

Comments

Labeling of GAPDH in pig TM cells using GAPDH antibody (2D4A7) at 1: 1000 dilution and Alexa 594 (red)

reviewed by:
Verified Customer

Human

09/19/2014

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Summary

Application	Western Blot
Sample Tested	GAPDH immunoblot in MCF7 cells
Species	Human

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reviewed by:
Bryan Tinsley

ICC

Human

06/10/2014

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Summary

Application	Immunocytochemistry
Species	Human

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reviewed by:
Bryan Tinsley

Human

06/10/2014

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Summary

Application	Western Blot
Species	Human

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reviewed by:
Bryan Tinsley

ICC

Human

05/21/2014

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Summary

Application	Immunocytochemistry
Species	Human

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reviewed by:
Mary Ndiaye

Human

06/10/2011

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Summary

Application	Western Blot
Sample Tested	HaCaT whole cell lysate, Sample Amount: 30 ug
Species	Human
Lot	B

Product General Protocols

View specific protocols for GAPDH Antibody (NB300-328):

Western Blot Protocol for GAPDH Antibody (NB300-328)

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FAQs for GAPDH Antibody (NB300-328). (Showing 1 - 7 of 7 FAQs).

We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
- 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 interested in. GAPDH antibody is a good choice for low to mid MW targets, as GAPDH detects around 35 kDa.
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.
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.
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.
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.
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.
I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
- For homogenized tissue, beta-actin and GAPDH are both fine.

Control Lysate(s)

GAPDH Overexpression Lysate

Secondary Antibodies

Isotype Controls

Additional GAPDH Products

Research Areas for GAPDH Antibody (NB300-328)

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Blogs on GAPDH. Showing 1-10 of 13 blog posts - Show all blog posts.

Check out the latest blog posts on GAPDH.

Considerations for Quantitative Western blotting
By Jamshed Arslan, Pharm. D., PhD. Since its inception in 1979, Western blotting has undergone several developments. The use of radioactive probes was common throughout 1980s, but utilizing secondary antibodies labell... Read full blog post.

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 western blots. However, this ... 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 Western Blot Loading Controls
What are Western Blot 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... 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.

Showing 1-10 of 13 blog posts - Show all blog posts.

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Recent Reviews

5	12
4	0
3	0
2	0
1	0

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	Verified Customer 04/04/2020
Application:	WB
Species:	Human

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	Verified Customer 12/27/2019
Application:	WB
Species:	Human

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	Verified Customer 02/19/2019
Application:	WB
Species:	Human and Mouse

read this review

Bioinformatics

Gene Symbol	GAPDH
Entrez	2597(Human) 14433(Mouse)
Uniprot	P04406(Human)

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

GAPDH Antibody (2D4A7)

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Publications for GAPDH Antibody (NB300-328)(68)

Reviews for GAPDH Antibody (NB300-328) (12) 512

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Product General Protocols

Video Protocols

FAQs for GAPDH Antibody (NB300-328). (Showing 1 - 7 of 7 FAQs).

Control Lysate(s)

Secondary Antibodies

Isotype Controls

Additional GAPDH Products

Research Areas for GAPDH Antibody (NB300-328)

Blogs on GAPDH. Showing 1-10 of 13 blog posts - Show all blog posts.

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Recent Reviews

Bioinformatics