GAPDH Antibody (1D4)

Western Blot: GAPDH Antibody (1D4) [NB300-221] - analysis of GAPDH in human nucleus pulposus cell culture (40ug/lane) using anti-GAPDH antibody. LI-COR Odyssey imaging system was used for detection. Image from verified customer review.

Western Blot: GAPDH Antibody (1D4) [NB300-221] - Analysis of cell line lysates probed with mouse mAb to GAPDH, NB300-221, dilution 1:2,000: [1] protein standard, [2] HEK293, [3] HeLa, [4] SH-SY5Y, [5] COS1, [6] NIH-3T3, and [7] C6 cells. The GAPDH antibody reveals a single band at ~37 kDa in all cell lines. GAPDH is a house keeping protein, the level of which is relatively unaffected by most experimental manipulations, and, as a result, this antibody has been widely used as a western blotting loading control.

Immunocytochemistry/Immunofluorescence: GAPDH Antibody (1D4) [NB300-221] - SH-SY5Y cells stained with GAPDH antibody NB300-221 (green) and Neurofilament Heavy antibody NB300-217 (red), counterstained with a fluorescent DNA probe (blue).

Western Blot: GAPDH Antibody (1D4) [NB300-221] - analysis of GAPDH in chicken pectoralis major using anti-GAPDH antibody. The primary antibody was used at a dilution of 1:4000, incubated for 1 hour at room temperature. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: GAPDH Antibody (1D4) [NB300-221] - SH-SY5Y cells stained with GAPDH antibody NB300-221 (green). Nuclear DNA is stained with Hoechst dye (blue).

• Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ch, Eq, Ha, Op, Pm, Sh, Ze
• Applications: WB, Simple Western, ChIP, ICC/IF, IHC, IP
• Clone: 1D4
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

GAPDH Antibody (1D4) Summary

• Description: This GAPDH antibody (1D4) is primarily used as a loading control in western blots due to the near global expression of GAPDH which can be attributed to its vital role in the glycolytic metabolic process.
• Immunogen: Purified porcine GAPDH [UniProt# P00355]
• Marker: Cytosolic Marker
• Isotype: IgM
• Clonality: Monoclonal
• Host: Mouse
• Gene: GAPDH
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000
  • Simple Western 1:25
  • Chromatin Immunoprecipitation 1:10-1:500
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunoprecipitation
• Application Notes: This GAPDH Antibody (1D4) is useful for Western blot and Immunocytochemistry/Immunofluorescence. Immunohistochemistry was reported in scientific literature. In WB a prominent band can be seen at approximately 38 kDa. This antibody makes an excellent loading control for Western blot experiments. Use in immunprecipitation reported in scientific literature (PMID 24032460). Use in chromatin immunoprecipitation reported in scientific literature (PMID 25802280).
  
  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: 37 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 (NBL1-10967)

Reactivity Notes

Human, mouse, rat, bovine, porcine, avian, sheep, opossum and primate. Likely to react with other mammals as well. Hamster and zebrafish reactivity reported by a customer review. chicken reactivity reported in scientific literature (PMID: 24725420)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% Glycerol
• Preservative: 5mM Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for GAPDH Antibody (1D4)

• 1D4 GAPDH
• aging-associated gene 9 protein
• Cytoplasm Marker
• EC 1.2.1
• EC 1.2.1.12
• EC 2.6.99.-
• G3PD antibody
• G3PD
• G3PDH
• GAPDH
• GAPDPeptidyl-cysteine S-nitrosylase GAPDH antibody
• GAPDPeptidyl-cysteine S-nitrosylase GAPDH
• glyceraldehyde 3-phosphate dehydrogenase
• glyceraldehyde-3-phosphate dehydrogenase
• MGC102544
• MGC102546
• MGC103190
• MGC103191
• MGC105239
• MGC127711
• MGC88685
• OCAS, p38
• OCT1 coactivator in S phase

Background

GAPDH is a critical enzyme in the glocolytic pathway, catalyzing the 6th step of glycolysis which is the beginning of the pay-off phase of glycolysis where a gain of ATP can be seen. GAPDH is primarily associated with metabolic functions but has also been shown to have a role in transcription activation, apoptosis and intracellular vesicle shuttling. Due to the importance of GAPDH in metabolic processes it is expressed in nearly all cells and therefore serves as a good loading control target. This GAPDH antibody, clone (1D4), is highly validated as a western blot loading control antibody and has been cited in over 180 publications. GAPDH, also known as Glyceraldehyde 3-Phosphate Dehydrogenase is a 146 kDa tetramer composed of four 30-40 kDa subunits and is reported to bind to a variety of other proteins, including the amyloid precursor protein. Associations with Actin and Tubulin have also been 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. This GAPDH antibody was used as a loading control in a study looking at the effects of RNAi treatment of HCV infected Huh-7 cells on expression levels of various proteins(1). This GAPDH antibody is cited as being used as a loading control in another study looking at the effects of mitogen-activated protein kinases on KChIP2 gene expression(2). The concentration of DPgly and DP in whole cytoplasm, Triton X-100 soluble cytoplasmic fractions and Triton-X 100 insoluble cytoplasmic fractions was analyzed in SCC9 cells at various stages of development; using GAPDH loading control antibody to account for differences in lysate loading(3). The effect of a p38 MAPK inhibitor (SB293063) vs vehicle treatment on the pKv2 phosphorylation state in cortical neurons was studied using this GAPDH antibody as a loading control to account for differences in lysate loading and overall expression levels(4). PMIDs: 1. 12566571: GAPDH Antibody Used as a Loading Control Antibody to Normalize Differences in Lysate Loading. 2. 16385079 Used GAPDH Antibody to Normalize Differences in Overall Expression Levels In Different Patients. 3. 16365169 GAPDH Antibody used as a Loading Control Antibody to Account for Discrepancies in SCC9 Lysate Loading. 4. 17360683 Considers the effect of a p38 MAPK inhibitor on the phosphorylation state of p38 MAPK and the progression of the apoptotic surge of potassium currents using the GAPDH loading control antibody to account for differences in lysate loading.

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.

Publications for GAPDH Antibody (NB300-221)(243)

Publications using NB300-221

• Chen L, Zhu G, Johns EM, Yang X. TRIM11 activates the proteasome and promotes overall protein degradation by regulating USP14. Nat Commun. Mar 26 2018 [PMID: 29581427] (WB, Human)
• Rudman M. Bromodomain and Extraterminal Domain-Containing Protein Inhibition Decreases Inflammation after Spinal Cord Injury Thesis 2017 (WB, Mouse)
• Troster P, Haseleu J, Petersen J, Drees O. The Absence of Sensory Axon Bifurcation Affects Nociception and Termination Fields of Afferents in the Spinal Cord. Front Mol Neurosci. 2018 Feb 08 [PMID: 29472841] (WB, Mouse)
• Fan CS, Chen WS, Chen LL, Chen CC. Osteopontin-integrin engagement induces HIF-1a-TCF12-mediated endothelial-mesenchymal transition to exacerbate colorectal cancer. Oncotarget. 2018 Jan 12 [PMID: 29435158] (WB, Human)
• Langhorst H, Juttner R, Groneberg D et al. The IgCAM CLMP is required for intestinal and ureteral smooth muscle contraction by regulating Connexin43 and 45 expression in mice Dis Model Mech 2018 Jan 19 [PMID: 29361518] (Mouse)
• Urata S, Izumi K, Hiratsuka K et al. C-C motif ligand 5 promotes migration of prostate cancer cells in the prostate cancer bone metastasis microenvironment. Cancer Sci. Mar 1 2018 [PMID: 29288523] (WB, Human)
• Fustaino V, Presutti D, Colombo T et al. Characterization of epithelial-mesenchymal transition intermediate/hybrid phenotypes associated to resistance to EGFR inhibitors in non-small cell lung cancer cell lines. Oncotarget 2017 Nov 28 [PMID: 29262566] (WB, Human)
• Lin H, Magrane J, Clark EM et al. Early VGLUT1-specific parallel fiber synaptic deficits and dysregulated cerebellar circuit in the KIKO mouse model of Friedreich ataxia. Dis Model Mech 2017 Dec 19 [PMID: 29259026] (WB, Mouse)
• Nguyen TP, Yong HE, Chollangi T et al. Altered downstream target gene expression of the placental Vitamin D receptor in human idiopathic fetal growth restriction Cell Cycle. 2017 Nov 22 [PMID: 29161966] (WB, Human)
• Liu C, Lee WC, Huang BM et al. 16-Hydroxycleroda-3, 13-dien-15, 16-olide inhibits the proliferation and induces mitochondrial-dependent apoptosis through Akt, mTOR, and MEK-ERK pathways in human renal carcinoma cells. Phytomedicine. 2017 Oct 03 [PMID: 29157834] (Human)
• Lin H, Magrane J, Rattelle A et al. Early cerebellar deficits in mitochondrial biogenesis and respiratory chain complexes in the KIKO mouse model of Friedreich ataxia Dis Model Mech. 2017 Nov 01 [PMID: 29125827] (WB, Mouse)
• Rostami J, Holmqvist S, Lindstrom V et al. Human astrocytes transfer aggregated alpha-synuclein via tunneling nanotubes J. Neurosci. 2017 Oct 31 [PMID: 29089438] (WB, Human)
• Han J, Plummer J, Liu L et al. The impact of obesity on brain iron levels and a-synuclein expression is regionally dependent. Nutr Neurosci. 2017 Oct 16 [PMID: 29034829] (WB, Mouse)
• Lin X, Wang L, Wang F. Chondromodulin-I suppresses tumorigenesis of human osteosarcoma cells. Mol Med Rep. 2017 Sep 26 [PMID: 28983591] (WB, Human)
• Sriwattanapong K, Slocum SL, Chawanthayatham S et al. Editor's Highlight: Pregnancy Alters Aflatoxin B1 Metabolism and Increases DNA Damage in Mouse Liver. Toxicol. Sci. Nov 1 2017 [PMID: 28973694] (WB, Mouse)
  
  Details:
  Novus GAPDH antibody clone 1D4 was used as a loading control for western blot to look at metabolic enzyme levels in pregnant vs nonpregnant mice
• Liao HF, Pan CH, Chou PY et al. Toxicological effects of NCKU-21, a phenanthrene derivative, on cell growth and migration of A549 and CL1-5 human lung adenocarcinoma cells PLoS ONE 2017 Sep 25 [PMID: 28945763] (WB, Human)
• Bove A, Gradeci D, Fujita Y et al. Local cellular neighbourhood controls proliferation in cell competition Mol. Biol. Cell 2017 Sep 20 [PMID: 28931601] (WB)
• Gillen AE, Brechbuhl HM, Yamamoto TM et al. Alternative Polyadenylation of PRELID1 Regulates Mitochondrial ROS Signaling and Cancer Outcomes Mol. Cancer Res. 2017 Sep 14 [PMID: 28912168] (WB, Human)
• Martin IV, Bohner A, Boor P et al. Complement C5a receptors C5L2 and C5aR in renal fibrosis Am. J. Physiol. Renal Physiol. 2017 Sep 13 [PMID: 28903945] (WB, Mouse)
• Xu Q, Jitkaew S, Choksi S et al. The cytoplasmic nuclear receptor RARg controls RIP1 initiated cell death when cIAP activity is inhibited Nat Commun 2017 Sep 04 [PMID: 28871172] (WB, Mouse)
• Vitte J, Gao F, Coppola G et al. Timing of Smarcb1 and Nf2 inactivation determines schwannoma versus rhabdoid tumor development Nat Commun 2017 Aug 21 [PMID: 28824165] (WB, Mouse)
• Zak TJ, Koshman YE, Samarel AM, Robia SL. Regulation of Focal Adhesion Kinase through a Direct Interaction with an Endogenous Inhibitor. Biochemistry 2017 Aug 23 [PMID: 28782937]
• George J, Tsutsumi M, Tsuchishima M. MMP-13 deletion decreases profibrogenic molecules and attenuates N-nitrosodimethylamine-induced liver injury and fibrosis in mice. J. Cell. Mol. Med. 2017 Aug 07 [PMID: 28782260] (WB, Mouse)
• Sui Z, Gokhin DS, Nowak RB et al. Stabilization of F-actin by tropomyosin isoforms regulates the morphology and mechanical behavior of red blood cells Mol. Biol. Cell 2017 Jul 18 [PMID: 28720661] (WB, Mouse)
• Lin NH, Messing A, Perng MD. Characterization of a panel of monoclonal antibodies recognizing specific epitopes on GFAP PLoS ONE 2017 Jul 12 [PMID: 28700643] (WB, Human)
• Sarowar T, Grabrucker S, Boeckers TM, Grabrucker, AM. Object Phobia and Altered RhoA Signaling in Amygdala of Mice Lacking RICH2. Front Mol Neurosci. 2017 Jun 23 [PMID: 28642683] (WB, Mouse)
• Al-Ali H, Ding Y, Slepak T et al. The mTOR substrate S6 Kinase 1 (S6K1) is a negative regulator of axon regeneration and a potential drug target for Central Nervous System injury. J. Neurosci. 2017 Jun 16 [PMID: 28626016] (WB, Rat)
• Chugh P, Clark AG, Smith MB et al. Actin cortex architecture regulates cell surface tension. Nat. Cell Biol. May 22 2017 [PMID: 28530659]
• Zhao N. Notch Signaling Guides Vascular Smooth Muscle Cell Function. Thesis. 2014 (WB, Human, Mouse)
• Almalki SG, Agrawal DK. ERK signaling is required for VEGF-A/VEGFR2-induced differentiation of porcine adipose-derived mesenchymal stem cells into endothelial cells. Stem Cell Res Ther May 12 2017 [PMID: 28499402] (WB, Porcine)
• Roux A, Loranger A, Lavoie JN, Marceau N. Keratin 8/18 regulation of insulin receptor signaling and trafficking in hepatocytes through a concerted phosphoinositide-dependent Akt and Rab5 modulation. FASEB J. Apr 25 2017 [PMID: 28442548] (WB)
• Chen HY, Chen CJ, Chen WC et al. A promising protein responsible for overactive bladder in ovariectomized mice. Taiwan J Obstet Gynecol. Apr 1 2017 [PMID: 28420508] (WB, Mouse)
• Ho TJ, Lee CW, Lu ZY et al. Effects of Electroacupuncture on Methamphetamine-Induced Behavioral Changes in Mice. Evid Based Complement Alternat Med 2017 Mar 16 [PMID: 28400844] (WB, Mouse)
• Du M, Martin A, Hays F et al. Serum retinol-binding protein-induced endothelial inflammation is mediated through the activation of toll-like receptor 4. Mol. Vis. Apr 12 2017 [PMID: 28400700] (WB, Human)
• Barr AR, Cooper S, Heldt FS et al. DNA damage during S-phase mediates the proliferation-quiescence decision in the subsequent G1 via p21 expression. Nat Commun. Mar 20 2017 [PMID: 28317845] (WB, Human)
• Abou-Kheir W Barrak J Hadadeh O, Daoud G. HTR-8/SVneo cell line contains a mixed population of cells. Placenta. [PMID: 28161053] (WB, Human)
• Sero JE, Bakal C. Multiparametric Analysis of Cell Shape Demonstrates that b-PIX Directly Couples YAP Activation to Extracellular Matrix Adhesion Cell Syst Jan 25 2017 [PMID: 28065575] (WB, Mouse)
• Maolake A, Izumi K, Shigehara K et al. Tumor-associated macrophages promote prostate cancer migration through activation of the CCL22-CCR4 axis. Oncotarget. Feb 7 2017 [PMID: 28039457] (WB, Human)
• Sankowski R, Herring A, Keyvani K et al. The multi-target effects of CNI-1493: convergence of anti-amylodogenic and anti-inflammatory properties in animal models of Alzheimer's disease. Mol. Med. Nov 15 2016 [PMID: 27847962] (WB, Mouse)
• Yoon YM, Storm KJ, Kamimae-Lanning AN et al. Endogenous DNA Damage Leads to p53-Independent Deficits in Replicative Fitness in Fetal Murine Fancd2(-/-) Hematopoietic Stem and Progenitor Cells. Stem Cell Reports Oct 6 2016 [PMID: 27720904] (WB, Mouse)
• Willi M, Yoo KH, Wang C et al. Differential cytokine sensitivities of STAT5-dependent enhancers rely on Stat5 autoregulation. Nucleic Acids Res. Sep 30 2016 [PMID: 27694626] (WB, Mouse)
• Drelon C, Berthon A, Sahut-Barnola I et al. PKA inhibits WNT signalling in adrenal cortex zonation and prevents malignant tumour development Nat Commun Sep 14 2016 [PMID: 27624192] (WB, Human)
• Sandin L, Bergkvist L, Nath S et al. Beneficial effects of increased lysozyme levels in Alzheimer's disease modelled in Drosophila melanogaster. FEBS J. Aug 26 2016 [PMID: 27562772] (WB)
• Pelham CJ, Drews EM, Agrawal DK. Vitamin D controls resistance artery function through regulation of perivascular adipose tissue hypoxia and inflammation. J Mol Cell Cardiol 2016 Sep [PMID: 27374117] (Mouse)
• Ochaba J, Monteys AM, O'Rourke JG et al. PIAS1 Regulates Mutant Huntingtin Accumulation and Huntington's Disease-Associated Phenotypes In Vivo. Neuron. 2016 May 04 [PMID: 27146268] (WB, Mouse)
• Ding L, Paszkowski-Rogacz M, Winzi M et al. Systems Analyses Reveal Shared and Diverse Attributes of Oct4 Regulation in Pluripotent Cells. Cell Syst 2015 Aug 26 [PMID: 27135800] (ICC/IF)
• Krapp C, Hotter D, Gawanbacht A et al. Guanylate Binding Protein (GBP) 5 Is an Interferon-Inducible Inhibitor of HIV-1 Infectivity. Cell Host Microbe. 2016 Mar 15 [PMID: 26996307] (WB, Human)
• Kumper S, Mardakheh FK, McCarthy A et al. Rho-associated kinase (ROCK) function is essential for cell cycle progression, senescence and tumorigenesis. Elife. 2016 Jan 14 [PMID: 26765561] (WB, Mouse)
  
  Details:
  GAPDH antibody used as a loading control for WB on mouse embryo fibroblasts (MEFs) from embryos with different genotypes (Figure 1F).
• Chen YC, Statt S, Wu R et al. High mobility group box 1-induced epithelial mesenchymal transition in human airway epithelial cells. Sci Rep. 2016 Jan 07 [PMID: 26739898] (WB, Human)
• Slocum SL, Skoko JJ, Wakabayashi N et al. Keap1/Nrf2 pathway activation leads to a repressed hepatic gluconeogenic and lipogenic program in mice on a high-fat diet. Arch. Biochem. Biophys. 2015 Dec 14 [PMID: 26701603] (WB, Mouse)
• Brai E, Marathe S, Astori S et al. Notch1 Regulates Hippocampal Plasticity Through Interaction with the Reelin Pathway, Glutamatergic Transmission and CREB Signaling. Front Cell Neurosci. 2015 Dec 04 [PMID: 26635527] (WB, Mouse)
• Gokhin DS, Ochala J, Domenighetti AA, Fowler VM. Tropomodulin1 directly controls thin filament length in both wild-type and tropomodulin4-deficient skeletal muscle. Development. 2015 Nov 19 [PMID: 26586224] (WB, Mouse)
• Presutti D, Santini S, Cardinali B et al. MET Gene Amplification and MET Receptor Activation Are Not Sufficient to Predict Efficacy of Combined MET and EGFR Inhibitors in EGFR TKI-Resistant NSCLC Cells. PLoS ONE. 2015 Nov 20 [PMID: 26580964] (WB, Human)
• Becs G, Zarjou A, Agarwal A et al. Pharmacological induction of ferritin prevents osteoblastic transformation of smooth muscle cells. J Cell Mol Med 2016 Feb [PMID: 26499096]
• DeLuca AP, Whitmore SS, Barnes J et al. Hypomorphic mutations in TRNT1 cause retinitis pigmentosa with erythrocytic microcytosis. Hum Mol Genet 2016 Jan 01 [PMID: 26494905] (WB)
• Asghar A, Lajeunesse A, Dulla K et al. Bub1 autophosphorylation feeds back to regulate kinetochore docking and promote localized substrate phosphorylation. Nat Commun 2015 Sep 24 [PMID: 26399325] (WB)
• Shang Y, Dong X, Han G et al. The Roles of Tight Junctions and Claudin-1 in the Microbubble-Mediated Ultrasound-Induced Enhancement of Drug Concentrations in Rat Prostate. J. Membr. Biol. 2015 Aug 20 [PMID: 26289600] (WB, Rat)
• Ikhapoh IA, Pelham CJ, Agrawal DK. Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells. Stem Cells Int 2015 [PMID: 26106428] (WB)
• Zakrzewicz A, Atanasova S, Padberg W. Monocytic Tissue Transglutaminase in a Rat Model for Reversible Acute Rejection and Chronic Renal Allograft Injury. Mediators of Inflammation. 2015 [PMID: 26063971] (WB, Rat)
• Boosani CS, Dhar K, Agrawal DK. Down-regulation of hsa-miR-1264 contributes to DNMT1-mediated silencing of SOCS3. Mol. Biol. Rep. 2015 Jun 06 [PMID: 26047583] (WB, Human)
  
  Details:
  GAPDH antibody (clone 1D4) used for WB as a loading control on HCASMCs (human coronary artery smooth muscle) cells. Cells were transfected with hsa-miR-1264 mimic or its inhibitor and treated with or without TNF-alpha and IGF-1 (Fig. 3).
• Serbo JV, Kuo S, Lewis S et al. Patterning of Fibroblast and Matrix Anisotropy within 3D Confinement is Driven by the Cytoskeleton Adv Healthc Mater. 2015 Jun 01 [PMID: 26033825]
• Chelluboina B, Warhekar A, Dillard M et al. Post-transcriptional inactivation of matrix metalloproteinase-12 after focal cerebral ischemia attenuates brain damage. Sci Rep. 2015 May 09 [PMID: 25955565] (WB, Rat)
• Baskar K, Sur S, Selvaraj V, Agrawal DK. Functional constituents of a local serotonergic system, intrinsic to the human coronary artery smooth muscle cells. Mol Biol Rep 2015 Aug [PMID: 25861735] (Human)
• Marathe S, Liu S, Brai E et al. Notch signaling in response to excitotoxicity induces neurodegeneration via erroneous cell cycle reentry Cell Death Differ. 2015 Mar 27 [PMID: 25822340] (IHC, ICC/IF, Mouse)
• Aboelenain M, Kawahara M, Balboula AZ et al. Status of autophagy, lysosome activity and apoptosis during corpus luteum regression in cattle J. Reprod. Dev. 2015 Mar 28 [PMID: 25819401] (WB, Bovine)
• Liu YN, Yin J, Barrett B et al. Loss of Androgen Regulated miR-1 Activates SRC and Promotes Prostate Cancer Bone Metastasis Mol. Cell. Biol. 2015 Mar 23 [PMID: 25802280] (WB, ChIP, Mouse)
• Zhou Q, Derti A, Ruddy DA et al. A chemical genetics approach for the functional assessment of novel cancer genes Cancer Res. 2015 Mar 18 [PMID: 25788694] (WB, Human)
• Aubry S, Shin W, Crary JF et al. Assembly and interrogation of Alzheimer's disease genetic networks reveal novel regulators of progression PLoS ONE 2015 Mar 18 [PMID: 25781952] (WB, Human)
• Bergert M, Erzberger A, Desai RA et al. Force transmission during adhesion-independent migration. Nat. Cell Biol. 2015 Apr 01 [PMID: 25774834] (WB, Human)
• Zakrzewicz A, Kutlimuratov K, Atanasova S et al. Attenuation of Acute Rat Renal Allograft Rejection by Apolipoprotein E-Mimetic Peptide Transplantation 2015 Feb 25 [PMID: 25719257] (WB, Human)
• De Boussac H, Maqdasy S, Trousson A et al. Enolase is regulated by Liver X Receptors Steroids. 2015 Feb 21 [PMID: 25708389] (WB, Mouse)
• Kutz JL, Greaney JL, Santhanam L, Alexander LM. Evidence for a functional vasodilatory role for hydrogen sulfide in the human cutaneous microvasculature J. Physiol. (Lond.). 2015 Jan 30 [PMID: 25639684] (WB, Human, Rat)
• Li J, Yuan W, Jiang S et al. Prolyl-4-hydroxylase Domain Protein 2 Controls NF-kB/p65 Transactivation and Enhances the Catabolic Effects of Inflammatory Cytokines on cells of the Nucleus Pulposus J. Biol. Chem. 2015 Jan 29 [PMID: 25635047] (WB, Rat, Human)
• Izuagie IA, Pelham CJ, Agrawal DK. Synergistic effect of angiotensin II on vascular endothelial growth factor-A-mediated differentiation of bone marrow-derived mesenchymal stem cells into endothelial cells. Stem Cell Res Ther 2015 Jan 6 [PMID: 25563650]
• Ye W, Zhou J, Markova DZ et al. Xylosyltransferase-1 Expression Is Refractory to Inhibition by the Inflammatory Cytokines Tumor Necrosis Factor alpha and IL-1b in Nucleus Pulposus Cells: Novel Regulation by AP-1, Sp1, and Sp3. Am. J. Pathol. 2014 Dec 02 [PMID: 25476526] (WB, Human)
  
  Details:
  GAPDH antibody used for WB on lysates of human nucleus pulposus (NP) cells transduced with LV-shC or LV-shSp1 or LV-shSp3 (Figure 6)
• Liu Jt, Bain Lj. Arsenic inhibits hedgehog signaling during P19 cell differentiation. Toxicol. Appl. Pharmacol. 2014 Dec 15 [PMID: 25448440] (WB, Mouse)
  
  Details:
  GAPDH antibody used for WB at 1:1000 dilution on lysates of mouse embryonal carcinoma P19 cell line which was induced to differentiate for 5 days while being exposed to 0, 0.25, or 0.5 uM arsenic/ sodium arsenite (Fig 3A and 3C; Fig 6).
• Chang Sh, Chang Wl, Lu Cc, Tarn Wy. Alanine repeats influence protein localization in splicing speckles and paraspeckles. Nucleic Acids Res. 2014 Nov 20 [PMID: 25414336]
• Zhao N, Koenig Sn, Trask Aj et al. mir145 Regulates TGFBR2 Expression and Matrix Synthesis in Vascular Smooth Muscle Cells. Circ. Res. 2014 Oct 16 [PMID: 25323858] (WB, Mouse)
• Peterson Jm, Mart R, Bond Ce. Effect of obesity and exercise on the expression of the novel myokines, Myonectin and Fibronectin type III domain containing 5. PeerJ. 2014 Oct 07 [PMID: 25289190] (WB, Rat)
• Domingue JC, Ao M, Sarathy J et al. HEK-293 cells expressing the cystic fibrosis transmembrane conductance regulator (CFTR): a model for studying regulation of Cl- transport. Physiol Rep. 2014 Sep 01 [PMID: 25263207] (WB, Human)
  
  Details:
  MRP4 antibody used for WB at 1:750 dilution on cytosolic and total lysate fractions of HEK?CFTR cells as well as T84 cells (positive control). The antibody detected a band of 170 kDa size and the data is shown in Figure 5A - lower panel.
• Chen X, Sadowska GB, Zhang et al. Neutralizing anti-interleukin-1b antibodies modulate fetal blood-brain barrier function after ischemia. Neurobiol. Dis. 2014 Sep 24 [PMID: 25258170] (WB, Sheep)
• Wang X, Wang H, Yang H et al. Tumor Necrosis Factor-a- and Interleukin-1b-Dependent Matrix Metalloproteinase-3 Expression in Nucleus Pulposus Cells Requires Cooperative Signaling via Syndecan 4 and Mitogen-Activated Protein Kinase-Nuclear Factor kB Axis: Implications in Inflammatory Disc Disease. Am. J. Pathol. 2014 Jul 22 [PMID: 25063530] (WB, Rat)
  
  Details:
  Glyceraldehyde-3-phosphate dehydrogenase / GAPDH antibody used for WB in TNF-alpha and IL-1 beta treated Rat's Nucleus pulposus/NP cells (Figure 1E, Figure 6A/E); TNF-alpha treated human NP cells transduced with either Sdc4-shRNA (LV-shSDC4) or Ctr-shRNA (LV-shC) (Figure 2C and E); human NP cells transduced with LV co-expressing YFP and either shp65 or shIkk beta (Figure 7C/E); TNF alpha/beta treated rat NP cells (Figure 8D).
• Palsamy P, Bidasee KR, Shinohara T. Selenite cataracts: Activation of endoplasmic reticulum stress and loss of Nrf2/Keap1-dependent stress protection. Biochim. Biophys. Acta 2014 Jul 02 [PMID: 24997453] (WB, Human)
  
  Details:
  GAPDH antibody used for WB on lysates of human lens epithelial cells (HLECs) treated or not with 10 uM sodium selenite for various time periods (Fig 6, Fig 7, Fig 8, Fig 11).
• Alidousty C, Rauen T, Hanssen L et al. Calcineurin-mediated YB-1 dephosphorylation regulates CCL5 expression during monocyte differentiation. J. Biol. Chem. 2014 Jun 19 [PMID: 24947514] (WB, Mouse, Rat, Human)
  
  Details:
  Used in WB of GAPDH to ensure equal protein loading of whole cell lysates or cytoplasmatic extracts from Human primary monocytes, THP-1 cells, HEK293 cells, rat mesangial cells (rMCs) and kidney of C57BL/6 mice.
• Holmes O, Paturi S, Selkoe DJ, Wolfe MS. Pen-2 Is Essential for gamma-Secretase Complex Stability and Trafficking but Partially Dispensable for Endoproteolysis Biochemistry. 2014 Jun 30 [PMID: 24941111] (WB, Mouse)
  
  Details:
  GAPDH antibody used as loading control in WB on lysates of Pen-2 knockout mouse embryonic fibroblasts/MEFs. Figure 6a - WB on stable cell lines of select Pen-2 mutants - monoclonal cell lines expressing wt, N8A, P27A, N33A, or I53A F-Pen-2; Figure 8 - WB on Pen-2 KO MEFs rescued with mutants of Pen-2 and treated with 8h - 10 uM MG132 ( a peptide aldehyde that effectively block the proteolytic activity of the 26S proteasome complex); Figure 10a - WB on Pen-2 KO MEFs treated with 10 uM MG132 or vehicle.
• McCord MC, Kullmann PH, He K et al. Syntaxin-binding domain of Kv2.1 is essential for the expression of apoptotic K+ currents. J. Physiol. (Lond.). 2014 Jun 13 [PMID: 24928958] (WB, Chinese Hamster)
  
  Details:
  Figure 3 C - Chinese hamster ovary (CHO) cells expressing syntaxin and Kv2.1 in the absence or presence of peptide C1a441–522
• Chelluboina B, Klopfenstein JD, Pinson DM et al. Stem Cell Treatment After Cerebral Ischemia Regulates the Gene Expression of Apoptotic Molecules Neurochem. Res. 2014 May 31 [PMID: 24879430] (WB, Rat)
  
  Details:
  GAPDH antibody used as a loading control for WB on the brain tissues of male Sprague–Dawley rats - sham controls and untreated or hUCBSCs/human umbilical cord blood-derived mesenchymal stem cells - treated ischemic brains (experimental MCAO/middle cerebral artery occlusion model intravenously injected or not with hUCBSCs). WB image in Fig. 4
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  Details:
  GAPDH (IMG-5019A-1). WB: Mouse embryonic P19 cells with/out exposure to arsenic, Figs 4D & 5C.
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  Details:
  WB: ATF6 (IMG-273) & GAPDH (IMG-5019A-1). Nuclear fractions of mouse uterus, Fig 6. Note: ATF6 (IMG-273) was seen at ~36 kDa, Fig 6.
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  Details:
  GAPDH-Loading Control (IMG-5019A-1). WB & IHC (frozen): P7 wild type and RGE -/- chicken retina, Fig 1B & Fig 1C.
• Swayne LA, Sorbara CD, Bennett SA. Pannexin 2 is expressed by postnatal hippocampal neural progenitors and modulates neuronal commitment. J Biol Chem. 2010 Aug 6 [PMID: 20529862] (WB)
  
  Details:
  WB: Neural stem and progenitor (NPC) cells isolated from postnatal day 3 C57BL/6 mouse hippocampus, Fig 1A. WB: Neuronal cells isolated from 14 day old C57BL/6 mouse neurospheres, Fig 2C & 3C.
• Hirai A, Ohtsuka N, Ikeda T et al. Role of mouse hepatitis virus (MHV) receptor murine CEACAM1 in the resistance of mice to MHV infection: studies of mice with chimeric mCEACAM1a and mCEACAM1b. J Virol. 2010 Jul [PMID: 20410265] (WB, Mouse)
  
  Details:
  WB: The liver, spleen, and ileum of MHV-A59 infected mice, Fig 3A.
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  Details:
  GAPDH (IMG-5019A-1): WB, human ocular pigment epithelial and normal aqueous cells, Fig 3.
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  Details:
  IP (human bronchial epithelial cells), Fig. 7A.
• Hynynen R, Suchanek M, Spl J et al. OSBP-related protein 2 is a sterol receptor on lipid droplets that regulates the metabolism of neutral lipids. J Lipid Res;50(7):1305-1315. 2009 [PMID: 19224871] (ICC/IF, Human)
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  Details:
  WB (human monocytes), Figs. 5A,C; 7A.
• Devalaraja-Narashimha K, Padanilam BJ. PARP-1 inhibits glycolysis in ischemic kidneys. J Am Soc Nephrol;20(1):95-103. 2009 Jan. [PMID: 19056868] (WB, Mouse)
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  Details:
  WB (human A549 and H460 cells), Fig. 1.
• Schmitt R, Marlier A, Cantley LG. Zag expression during aging suppresses proliferation after kidney injury. J Am Soc Nephrol;19(12):2375-83. 2008 Dec. [PMID: 18815245] (WB, Mouse)
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  Details:
  WB (mouse hippocampal lysates), Fig. 7A.
• Macartney-Coxson DP, Hood KA, Shi HJ et al. Metastatic susceptibility locus, an 8p hot-spot for tumour progression disrupted in colorectal liver metastases: 13 candidate genes examined at the DNA, mRNA and protein level. BMC Cancer. 2008 Jul 1 [PMID: 18590575] (WB, Human)
  
  Details:
  GAPDH (IMG-5019A). WB: Human colon normal, colon tumour, and liver metastasis, Fig 5.
• Turnquist HR et al. IL-1beta-driven ST2L expression promotes maturation resistance in rapamycin-conditioned dendritic cells. J Immunol;181(1):62-72. 2008 Jul 1. [PMID: 18566370] (WB, Mouse)
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• Diaz Encarnacion MM et al. Signaling pathways modulated by fish oil in salt-sensitive hypertension. Am J Physiol Renal Physiol;294(6):F1323-35. 2008 Jun. [PMID: 18385269] (WB, Rat)
• Zhang QS et al. Tempol protects against oxidative damage and delays epithelial tumor onset in Fanconi anemia mice. Cancer Res;68(5):1601-8. 2008 Mar 1. [PMID: 18316625] (WB, Mouse)
• Chen, K, Y Tu et al. PINCH-1 regulates the ERK-Bim pathway and contributes to apoptosis resistance in cancer cells. J Biol Chem. 283(5):2508-17. 2008 Feb 1. [PMID: 18063582] (WB, Human)
• Tsai WJ, McCormick KM, Brazeau DA et al. Estrogen effects on skeletal muscle insulin-like growth factor 1 and myostatin in ovariectomized rats. Exp Biol Med (Maywood). 2007 Nov [PMID: 17959844] (WB, Rat)
  
  Details:
  WB (used as a gel loading control for rat muscle tissue lysates): Figs 6C, 7C, 8C.
• Limes, S W, P J Rozance et al. Increased insulin sensitivity and maintenance of glucose utilization rates in fetal sheep with placental insufficiency and intrauterine growth restriction. Am J Physiol Endocrinol Metab;293(6):E1716-25. 2007 Dec. [PMID: 17895285] (WB, Sheep)
• Banerjee PP, Pandey R, Zheng R et al. Cdc42-interacting protein-4 functionally links actin and microtubule networks at the cytolytic NK cell immunological synapse. J Exp Med. 2007 Oct 1 [PMID: 17785506] (WB)
  
  Details:
  WB (Fig 8A): Protein standardization for siRNA studies.
• Cui, W, E Cuartas et al. CD200 and its receptor, CD200R, modulate bone mass via the differentiation of osteoclasts. Proc Natl Acad Sci U S A;104(36):14436-41. 2007 Sep 4. [PMID: 17726108] (WB, Mouse)
• Woeller CF, Fox JT, Perry C, Stover PJ. A ferritin-responsive internal ribosome entry site regulates folate metabolism. J Biol Chem;282(41):29927-35. 2007 Oct 12. [PMID: 17702748] (WB, Human)
• Porchia, LM et al. 5-(2-phenanthrenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]-phenyl} acetamide (OSU-03012), a celecoxib derivative, directly targets p21-activated kinase. Mol Pharmacol. 72(5):1124-31. Epub 2007 Aug 2. 2007 Nov. [PMID: 17673571] (WB, Human)
• Zhou M, Xia L, Wang J. Metformin transport by a newly cloned proton-stimulated organic cation transporter (plasma membrane monoamine transporter) expressed in human intestine. Drug Metab Dispos. 2007 Oct [PMID: 17600084] (WB, Human)
  
  Details:
  WB (human small intestine, Fig 6B): 1. GAPDH (IMG-5019A-1, -2) 2. INSTA-BLOT containing human small intenstine tissue lysate (IMB-103, IMB-123).
• Mao W, Fukuoka S, Iwai C et al. Cardiomyocyte apoptosis in autoimmune cardiomyopathy: mediated via endoplasmic reticulum stress and exaggerated by norepinephrine. Am J Physiol Heart Circ Physiol. 2007 Sep [PMID: 17545481] (WB, Rabbit)
  
  Details:
  Antibodies cited: 1. ATF6 (IMG-273): WB (Fig 7), analysis of ATF6 in nuclear fraction of rabbit left ventricle myocardium tissue lysates, Authors describe cleaved forms of p90ATF6 in the nucleus: p60ATF6 and p36ATF6. 2. GAPDH (IMG-5019A): WB (Fig 4), GAPD
• Blight KJ. Allelic variation in the hepatitis C virus NS4B protein dramatically influences RNA replication. J Virol. 2007 Jun [PMID: 17360748] (WB, Human)
  
  Details:
  Antibodies cited: 1. GAPDH (IMG-5019A-1, IMG-5019A-2) [WB, Fig7, Fig8 (Huh-7.5 human hepatoma cell line)].
• Redman, P T et al. Apoptotic surge of potassium currents is mediated by p38 phosphorylation of Kv2.1. Proc Natl Acad Sci U S A. 104(9):3568-73. 2007 Feb 27. [PMID: 17360683] (WB, Rat)
• Block, K et al. NAD(P)H oxidases regulate HIF-2alpha protein expression. J Biol Chem;282(11):8019-26. 2007 Mar 16. [PMID: 17200123] (WB, Human)
• Xia L, Engel K, Zhou M et al. Membrane localization and pH-dependent transport of a newly cloned organic cation transporter (PMAT) in kidney cells. Am J Physiol Renal Physiol. 2007 Feb [PMID: 17018840]
• Wang G, Wu WW, Pisitkun T et al. Automated quantification tool for high-throughput proteomics using stable isotope labeling and LC-MSn. Anal Chem 2006 Aug 15 [PMID: 16906720]
• Fatma S, Yakubov R, Anwar K, Hussain MM. Pluronic L81 enhances triacylglycerol accumulation in the cytosol and inhibits chylomicron secretion. J Lipid Res;47(11):2422-32. 2006 Nov. [PMID: 16894241] (WB, Human)
• Ju, J-S et al. Potentiation of insulin-stimulated glucose transport by the AMP-activated protein kinase. Am J Physiol Cell Physiol;292(1):C564-72. 2007 Jan. [PMID: 16870829] (WB, Mouse)
• Kanadia, R N et al. Reversal of RNA missplicing and myotonia after muscleblind overexpression in a mouse poly(CUG) model for myotonic dystrophy. Proc Natl Acad Sci U S A;103(31):11748-53. 2006 Aug 1. [PMID: 16864772] (WB, Mouse)
• Iwatani M, Ikegami K, Kremenska Y et al. Dimethyl sulfoxide has an impact on epigenetic profile in mouse embryoid body. Stem Cells. 2006 Nov [PMID: 16840553]
• Shibata, Yoshimi et al. Heat-killed BCG induces biphasic cyclooxygenase 2+ splenic macrophage formation--role of IL-10 and bone marrow precursors. J Leukoc Biol;80(3):590-8. 2006 Sep. [PMID: 16822852] (WB, Mouse)
• Khan N, Gordon R, Woodruff TM. Antiallodynic effects of alpha lipoic acid in an optimized RR-EAE mouse model of MS-neuropathic pain are accompanied by attenuation of upregulated BDNF-TrkB-ERK signaling in the dorsal horn of the spinal cord. Pharmacology Research & Perspectives. 2015 (WB, Mouse)
• Le Boeuf F, Houle F, Sussman M et al. Phosphorylation of focal adhesion kinase (FAK) on Ser732 is induced by rho-dependent kinase and is essential for proline-rich tyrosine kinase-2-mediated phosphorylation of FAK on Tyr407 in response to vascular endothelial growth factor. Mol Biol Cell;17(8):3508-20. 2006 Aug. [PMID: 16760434] (WB, Human)
• Barry, JS et al. Developmental changes in ovine myocardial glucose transporters and insulin signaling following hyperthermia-induced intrauterine fetal growth restriction. Exp Biol Med (Maywood);231(5):566-75. 2006 May. [PMID: 16636305] (WB, Sheep)
• Azfer, A et al. Activation of endoplasmic reticulum stress response during the development of ischemic heart disease. Am J Physiol Heart Circ Physiol;291(3):H1411-20. 2006 Sep. [PMID: 16617122] (WB, Mouse)
• Arthur DB et al. Inhibition of apoptosis by P2Y2 receptor activation: novel pathways for neuronal survival. J Neurosci;26(14):3798-804. 2006 Apr 5. [PMID: 16597733] (WB, Mouse)
• Pisitkun, T et al. High-throughput identification of IMCD proteins using LC-MS/MS. Physiol Genomics;25(2):263-76. 2006 Apr 13. [PMID: 16449382] (WB, Rat)
• Jia, Y Takimoto, K Jia Y, Takimoto K. Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res;98(3):386-93. 2006 Feb 17. [PMID: 16385079] (WB, Rat)
• Godsel, LM et al. Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol;171(6):1045-59. 2005 Dec 19. [PMID: 16365169] (WB, Human)
• Dusek, R L et al. The differentiation-dependent desmosomal cadherin desmoglein 1 is a novel caspase-3 target that regulates apoptosis in keratinocytes. J Biol Chem;281(6):3614-24. 2006 Feb 10. [PMID: 16286477] (WB, Human)
• Liu X, Kvetnansky R, Serova L et al. Increased susceptibility to transcriptional changes with novel stressor in adrenal medulla of rats exposed to prolonged cold stress. Brain Res Mol Brain Res. 2005 Nov 18 [PMID: 16169632]
• Pendelton, LC et al. Regulation of endothelial argininosuccinate synthase expression and NO production by an upstream open reading frame. J Biol Chem;280(25):24252-60. 2005 Jun 24. [PMID: 15851478] (WB, Bovine)
• Sarkar, S et al. Myocardial cell death and regeneration during progression of cardiac hypertrophy to heart failure. J Biol Chem;279(50):52630-42. 2004 Dec 10. [PMID: 15385543] (WB, Rat, Human)
• Gonzales-Cabrera, P et al. Differential regulation of the cell cycle by alpha1-adrenergic receptor subtypes. Endocrinology;145(11):5157-67. 2004 Nov. [PMID: 15297446] (WB, Rat, Human)
• Moshynska, O et al. Prognostic significance of a short sequence insertion in the MCL-1 promoter in chronic lymphocytic leukemia. J Natl Cancer Inst;96(9):673-82. 2004 May 5. [PMID: 15126604] (WB, Human)
• Akassoglou, K et al. Brain-specific deletion of neuropathy target esterase/swisscheese results in neurodegeneration. Proc Natl Acad Sci U S A;101(14):5075-80. 2004 Apr 6. [PMID: 15051870] (WB, IHC, ICC/IF, Mouse)
• Ellis RC, Earnhardt JN, Hayes RL et al. Cathepsin B mRNA and protein expression following contusion spinal cord injury in rats. J Neurochem. 2004 Feb [PMID: 14720218] (WB)
  
  Details:
  WB: GAPDH used as a protein loading control (rat spinal cord tissue), Fig. 3a,b,c.
• Fortun J, Dunn WA Jr, Joy S et al. Emerging role for autophagy in the removal of aggresomes in Schwann cells. J Neurosci. 2003 Nov 19 [PMID: 14627652] (WB, Mouse)
  
  Details:
  WB: GAPDH used as a protein loading control (Mouse Schwann cell lysates), Fig. 3F.
• Kapadia, SB et al. Kapadia SB, Brideau-Andersen A, Chisari FV. Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc Natl Acad Sci U S A;100(4):2014-8. 2003 Feb 18. [PMID: 12566571] (WB, ICC/IF, Human)

Reviews for GAPDH Antibody (NB300-221) (16)

Average Rating: 4.3

(Based on 16 reviews)

We have 16 reviews tested in 5 species: Human, Mouse, Rat, Mouse and Human, Other.

reviewed by: Anonymous WB Mouse and Human 06/29/2017

Summary

• Application: Western Blot
• Sample Tested: 293FT cell lysate,nih3t3 cell lysate
• Species: Mouse and Human
• Lot: 120116

reviewed by: Jim Hsiao WB Mouse 05/22/2017

Summary

• Application: Western Blot
• Sample Tested: Mouse lung endothelial cells
• Species: Mouse
• Lot: 012517

Comments

• Comments: Cell lysates of treated mouse lung endothelial cells were analyzed by western blotting.

reviewed by: Anonymous WB Human 01/14/2015

Summary

• Application: Western Blot
• Sample Tested: Human nucleus pulposus cell culture
• Species: Human
• Lot: 111711

Blocking

• Blocking Details: Li-Cor WB blocking buffer at 1X for 1h at room temperature.

Primary Anitbody

• Dilution Ratio: 1:500 dilution. Incubate overnight at 4 degree with gently shaking. Buffer: 50% of Li-cor blocking buffer plus 50% of PBS.

Secondary Antibody

• Secondary Description: GOAT-anti-mouse IRDye 680
• Secondary Manufacturer Cat#: Li-Cor #926-32220
• Secondary Concentration: 1:10000

Details

• Detection Notes: Li-Cor Odyssey. Membrane scan mode. postive control is A431 cell lysate. negative control is untreated cells.

Comments

• Comments: This is product works similar to the other GAPDH we had in this experiment. Backgroud is very little.

reviewed by: Anonymous WB Other 01/09/2015

Summary

• Application: Western Blot
• Sample Tested: Chicken Pectoralis Major
• Species: Other
• Lot: 111711

Blocking

• Blocking Details: 3% Casein in PBS with 0.1% Tween-20 for 1 hr

Primary Anitbody

• Dilution Ratio: 1/4000 at Room Temperature for 1 hr

Secondary Antibody

• Secondary Description: Goat anti-Mouse 680 RD - Licor Biosciences
• Secondary Manufacturer Cat#: 926-68180
• Secondary Concentration: 1/25,000

Details

• Detection Notes: Licor Odyssey

reviewed by: Eric England WB Other 10/08/2014

Summary

• Application: Western Blot
• Sample Tested: Whole Skeletal Muscle Lysate
• Species: Other
• Lot: 111711

Blocking

• Blocking Details: 3% Casein in PBS with 0.1% Tween-20 for 1 hr

Primary Anitbody

• Dilution Ratio: 1/2000 at 4C for 18 hr

Secondary Antibody

• Secondary Description: Goat anti-Mouse 680 RD - Licor Biosciences
• Secondary Manufacturer Cat#: 926-68180
• Secondary Concentration: 1/25,000

Details

• Detection Notes: Licor Odyssey

reviewed by: Liyong Zhang WB Mouse 07/01/2014

Summary

• Application: Western Blot
• Species: Mouse
• File: View PDF

reviewed by: Anonymous WB Other 12/16/2013

Summary

• Application: Western Blot
• Sample Tested: Zebrafish embryo lysates
• Species: Other
• Comments: Zebrafish embryo lysates at different developmental stages probed with GAPDH antibody 1D4.

Comments

• Comments: Zebrafish embryo lysates at different developmental stages probed with GAPDH antibody 1D4.

reviewed by: Anonymous WB Human 12/23/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:2000, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: Anti-Mouse HRP

Details

• Detection Notes: Detection Method: ECL

reviewed by: Anonymous WB Human 12/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human Hepatoblastoma
• Species: Human
• Comments: The cultured cells were harvested, washed and lysed. Extracts were stored at -20C prior to analysis

Blocking

• Blocking Details: Blocking Buffer: 5% non-fat milk in PBST, Blocking Time: 1 hour

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: 24 hours, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab: Goat anti-Mouse IgG (HRP), Jackson

Details

• Detection Notes: Detection Method: GE ImageQuant LAS 4000 mini plus ECL

Comments

• Comments: The cultured cells were harvested, washed and lysed. Extracts were stored at -20C prior to analysis

reviewed by: Anonymous WB Rat 12/06/2011

Summary

• Application: Western Blot
• Sample Tested: Rat
• Species: Rat

Blocking

• Blocking Details: Blocking Buffer: 5% Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: 5 hours

Secondary Antibody

• Secondary Description: Secondary Ab: Mouse

Details

• Detection Notes: Detection Method: ECL

reviewed by: Anonymous WB Human 12/05/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab Dilution Ratio: 1:5000

Details

• Detection Notes: Detection Method: ECL

reviewed by: Anonymous WB Rat 11/23/2011

Summary

• Application: Western Blot
• Sample Tested: Rat brain
• Species: Rat
• Comments: Good!

Blocking

• Blocking Details: Blocking Buffer: 1% milk, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1500, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab: anti-mouse, Secondary Ab Dilution Ratio: 1:2000

Details

• Detection Notes: Detection Method: ECL

Comments

• Comments: Good!

reviewed by: Anonymous WB Rat 11/21/2011

Summary

• Application: Western Blot
• Sample Tested: Rat
• Species: Rat
• Comments: ok

Blocking

• Blocking Details: Blocking Buffer: 5% non-fat milk in TBST

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: AP-conjugate antibody

Comments

• Comments: ok

reviewed by: Anonymous WB Human 09/30/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human
• Comments: Very good!

Blocking

• Blocking Details: Blocking Buffer: 5% milk in 0.1% PBST

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:5000 diluted in blocking solution, Primary Ab Incubation Time: overnight, Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab Dilution Ratio: 1:5000 diluted in blocking solution

Details

• Detection Notes: Detection Method: ECL

Comments

• Comments: Very good!

reviewed by: Anonymous WB Other 03/25/2010

Summary

• Application: Western Blot
• Sample Tested: CHO cells, Sample Amount: 20ug
• Species: Other
• Lot: 113003

Blocking

• Blocking Details: Blocking Buffer: 5% milk TBS/T, Blocking Time: 1 hour, Blocking Temp: 20 degrees Celsius

Primary Anitbody

• Dilution Ratio: Primary Ab Incubation Dilution Buffer: 5% milk TBS/T, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab: goat anti-mouse mAb, conj. HRP

reviewed by: Anonymous WB Human 06/04/2009

Summary

• Application: Western Blot
• Sample Tested: EAhy926 cell lysate, Sample Amount: 15ug
• Species: Human
• Lot: P1
• Comments: We've been using this product for many years.

Blocking

• Blocking Details: Blocking Buffer: StartingBlock, Blocking Time: 2 hours, Blocking Temp: 25 degrees Celsius

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1/5000, Incubation Dilution Buffer: 1:5 StartingBlock to Water, Incubation Time: overnight, Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab: anti-mouse, Secondary Ab Dilution Ratio: 1/50000

Details

• Detection Notes: Detection Method: ECL, Exposure Time: 2 seconds, Specific Bands: 30-40 kDa

Comments

• Comments: We've been using this product for many years.

FAQs for GAPDH Antibody (NB300-221). (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.

Bioinformatics

• Gene Symbol: GAPDH
• Entrez:
  • 2597(Human)
  • 14433(Mouse)
  • 24383(Rat)
• Uniprot:
  • P00355(Porcine)