Skip to content

rabbit anti-GAPDH polyclonal antibody 1574

$100.00$2,600.00

Antibody summary

  • Rabbit polyclonal to GAPDH
  • Suitable for: WB, ICC/IF, IHC
  • Reacts with: human, mouse, rat
  • Isotype: IgG
  • 100 µL, 25 µL, 1 mL
SKU: 1574parent Categories: , Tag:
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

serum

applications

ICC/IF, IHC, WB

reactivity

human, mouse, rat

available sizes

1 mg, 100 µg, 25 µg

rabbit anti-GAPDH polyclonal antibody 1574

antibody
Database link:
human P04406
mouse P16858
rat P04797
Tested applications
WB,IHC,IHC,ICC/IF
Recommended dilutions
WB: 1:30000 IF/ICC and IHC: 1:2000
Immunogen
Full length recombinant human GAPDH expressed in and purified from E. coli
Size and concentration
25, 100, 1000µL and serum
Form
liquid
Storage Instructions
2-8°C for short term, for longer term at -20°C. Avoid freeze / thaw cycles.
Storage buffer
serum, 0.04% NaN3 added
Purity
serum
Clonality
polyclonal
Isotype
IgG
Compatible secondaries
goat anti-rabbit IgG, H&L chain specific, peroxidase conjugated, conjugated polyclonal antibody 9512
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody 2079
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody 7863
goat anti-rabbit IgG, H&L chain specific, Cross Absorbed polyclonal antibody 2371
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1715
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1720
Isotype control
Rabbit polyclonal - Isotype Control
target relevance
GAPDH plays a crucial role in glycolysis, the metabolic pathway responsible for generating energy in cells. Due to its stable expression levels and essential role in cellular metabolism, GAPDH is commonly used as a loading control in various experimental techniques, such as Western blotting. As a loading control, GAPDH serves as a reference protein to normalize and ensure equal loading of protein samples across different lanes on a gel, compensating for variations in protein loading and gel transfer. Its relatively constant expression in most cells and tissues makes it an ideal standard for comparing protein levels, allowing researchers to accurately quantify and compare target protein expression in experimental samples. This antibody can be used as a loading control when run alongside proteins of interest with different and resolvable molecular weights and ideally in combination with antibodies of same host and when using a secondary antibody.

Click for more on: loading controls and GAPDH
Protein names
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EC 1.2.1.12) (Peptidyl-cysteine S-nitrosylase GAPDH) (EC 2.6.99.-)
Gene names
GAPDH,GAPDH GAPD CDABP0047 OK/SW-cl.12
Protein family
Glyceraldehyde-3-phosphate dehydrogenase family
Mass
36053Da
Function
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively (PubMed:3170585, PubMed:11724794). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate (PubMed:3170585, PubMed:11724794). Modulates the organization and assembly of the cytoskeleton (By similarity). Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes (PubMed:23071094). Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation (PubMed:23071094). Also plays a role in innate immunity by promoting TNF-induced NF-kappa-B activation and type I interferon production, via interaction with TRAF2 and TRAF3, respectively (PubMed:23332158, PubMed:27387501). Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis (By similarity). Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity).
Catalytic activity
Reaction=D-glyceraldehyde 3-phosphate + NAD(+) + phosphate = (2R)-3-phospho-glyceroyl phosphate + H(+) + NADH; Xref=Rhea:RHEA:10300, ChEBI:CHEBI:15378, ChEBI:CHEBI:43474, ChEBI:CHEBI:57540, ChEBI:CHEBI:57604, ChEBI:CHEBI:57945, ChEBI:CHEBI:59776; EC=1.2.1.12; Evidence=; Reaction=L-cysteinyl-[protein] + S-nitroso-L-cysteinyl-[GAPDH] = L-cysteinyl-[GAPDH] + S-nitroso-L-cysteinyl-[protein]; Xref=Rhea:RHEA:66684, Rhea:RHEA-COMP:10131, Rhea:RHEA-COMP:17089, Rhea:RHEA-COMP:17090, Rhea:RHEA-COMP:17091, ChEBI:CHEBI:29950, ChEBI:CHEBI:149494; Evidence=; PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66685; Evidence=;
Pathway
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
Subellular location
Cytoplasm, cytosol Nucleus Cytoplasm, perinuclear region Membrane Cytoplasm, cytoskeleton Note=Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions (PubMed:12829261).
Structure
Homotetramer (PubMed:16239728, PubMed:16510976). Interacts with TPPP; the interaction is direct (By similarity). Interacts (when S-nitrosylated) with SIAH1; leading to nuclear translocation (By similarity). Interacts with RILPL1/GOSPEL, leading to prevent the interaction between GAPDH and SIAH1 and prevent nuclear translocation (By similarity). Interacts with CHP1; the interaction increases the binding of CHP1 with microtubules (By similarity). Associates with microtubules (By similarity). Interacts with EIF1AD, USP25, PRKCI and WARS1 (PubMed:11724794, PubMed:16501887, PubMed:15628863, PubMed:20644585). Interacts with phosphorylated RPL13A; inhibited by oxidatively-modified low-densitity lipoprotein (LDL(ox)) (PubMed:22771119). Component of the GAIT complex (PubMed:15479637). Interacts with FKBP6; leading to inhibit GAPDH catalytic activity (PubMed:19001379). Interacts with TRAF2, promoting TRAF2 ubiquitination (PubMed:23332158). Interacts with TRAF3, promoting TRAF3 ubiquitination (PubMed:27387501).
Post-translational modification
S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus (By similarity). S-nitrosylation of Cys-247 is induced by interferon-gamma and LDL(ox) implicating the iNOS-S100A8/9 transnitrosylase complex and seems to prevent interaction with phosphorylated RPL13A and to interfere with GAIT complex activity (PubMed:22771119, PubMed:25417112). ; ISGylated. ; Sulfhydration at Cys-152 increases catalytic activity. ; Oxidative stress can promote the formation of high molecular weight disulfide-linked GAPDH aggregates, through a process called nucleocytoplasmic coagulation. Such aggregates can be observed in vivo in the affected tissues of patients with Alzheimer disease or alcoholic liver cirrhosis, or in cell cultures during necrosis. Oxidation at Met-46 may play a pivotal role in the formation of these insoluble structures. This modification has been detected in vitro following treatment with free radical donor (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide. It has been proposed to destabilize nearby residues, increasing the likelihood of secondary oxidative damages, including oxidation of Tyr-45 and Met-105. This cascade of oxidations may augment GAPDH misfolding, leading to intermolecular disulfide cross-linking and aggregation. ; Succination of Cys-152 and Cys-247 by the Krebs cycle intermediate fumarate, which leads to S-(2-succinyl)cysteine residues, inhibits glyceraldehyde-3-phosphate dehydrogenase activity. Fumarate concentration as well as succination of cysteine residues in GAPDH is significantly increased in muscle of diabetic mammals. It was proposed that the S-(2-succinyl)cysteine chemical modification may be a useful biomarker of mitochondrial and oxidative stress in diabetes and that succination of GAPDH and other thiol proteins by fumarate may contribute to the metabolic changes underlying the development of diabetes complications. ; (Microbial infection) Glycosylated by C.rodentium protein NleB, enteropathogenic E.coli protein NleB1 and S.typhimurium protein Ssek1: arginine GlcNAcylation prevents the interaction with TRAF2 and TRAF3 (PubMed:23332158, PubMed:27387501, PubMed:28522607). This leads to reduced ubiquitination of TRAF2 and TRAF3, and subsequent inhibition of NF-kappa-B signaling and type I interferon production, respectively (PubMed:23332158, PubMed:27387501).
Target Relevance information above includes information from UniProt accession: P04406
The UniProt Consortium

ICC/IF-image-rabbit-anti-GAPDH-polyclonal-antibody-1574
Western blot analysis of equal amounts of cell lysates using mouse mAb to Ki67, 7671, dilution 1:2,000, in green: [1] protein standard (red), [2] rapidly dividing HeLa cell cultures, [3] rapidly dividing HEK293 cell cultures. Strong double bands above 250kDa correspond to the two major Ki67 isoforms of apparent molecular weight of 345 and 395kDa. Smaller proteolytic fragments of these isoforms are also invariably detected on the blot.
IHC-image-rabbit-anti-GAPDH-polyclonal-antibody-1574
Chromogenic immunostaining of a NBF fixed paraffin embedded human pancreas section with rabbit pAb to GAPDH, 1574, dilution 1:5,000, detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. The GAPDH antibody labels pancreatic exocrine glandular cells as well as a sub-population of endocrine cells. This antibody performs well in testing with both 4% PFA and standard NBF fixed human, mouse, and rat tissues.
WB-image-rabbit-anti-GAPDH-polyclonal-antibody-1574
Western blot analysis of different cell cytosolic or nuclear enriched fractions, using rabbit pAb to GAPDH, 1574, dilution 1:20,000, in red: [1] protein standard, [2] NIH-3T3 cytosolic, [3] NIH-3T3 nuclear, [4] HeLa cytosolic, and [5] HeLa nuclear fractions. Strong band at 37kDa corresponds to GAPDH protein, mainly detected in the cytosolic fractions. The same blot was simultaneously probed with mouse mAb to the nuclear RNA binding protein SF3B4 dilution 1:1,000, in green. In contrast to GAPDH, SF3B4 is exclusively expressed in the nuclear fraction.

Publications

pmidtitleauthorscitation
36565814High fat diet exacerbates long-term metabolic, neuropathological, and behavioral derangements in an experimental mouse model of traumatic brain injuryIbeh S, Bakkar NZ, Ahmad F, Nwaiwu J, Barsa C, Mekhjian S, Reslan MA, Eid AH, Harati H, Nabha S, Mechref Y, El-Yazbi AF, Kobeissy F.Life Sci. 2023 Feb 1;314:121316. doi: 10.1016/j.lfs.2022.121316. Epub 2022 Dec 22.
34685547Zonula occludens 2 and Cell-Cell Contacts Are Required for Normal Nuclear Shape in EpitheliaHernández-Guzmán C, Gallego-Gutiérrez H, Chávez-Munguía B, Martín-Tapia D, González-Mariscal L.Cells. 2021 Sep 28;10(10):2568. doi: 10.3390/cells10102568.
32945372E7 oncoprotein from human papillomavirus 16 alters claudins expression and the sealing of epithelial tight junctionsUc PY, Miranda J, Raya-Sandino A, Alarcón L, Roldán ML, Ocadiz-Delgado R, Cortés-Malagón EM, Chávez-Munguía B, Ramírez G, Asomoza R, Shoshani L, Gariglio P, González-Mariscal L.Int J Oncol. 2020 Oct;57(4):905-924. doi: 10.3892/ijo.2020.5105. Epub 2020 Jul 29.
31017387HSP90 Inhibitor, NVP-AUY922, Improves Myelination in Vitro and Supports the Maintenance of Myelinated Axons in Neuropathic MiceChittoor-Vinod VG, Bazick H, Todd AG, Falk D, Morelli KH, Burgess RW, Foster TC, Notterpek L.ACS Chem Neurosci. 2019 Jun 19;10(6):2890-2902. doi: 10.1021/acschemneuro.9b00105. Epub 2019 May 3.
30209366GAPDH inhibits intracellular pathways during starvation for cellular energy homeostasisYang JS, Hsu JW, Park SY, Li J, Oldham WM, Beznoussenko GV, Mironov AA, Loscalzo J, Hsu VW.Nature. 2018 Sep;561(7722):263-267. doi: 10.1038/s41586-018-0475-6. Epub 2018 Sep 12.
Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.

relevant to this product
Western blot
IHC
ICC
#
Please enter your product and batch number here to retrieve - product datasheet, SDS, and QC information.

Reviews

There are no reviews yet.

Only logged in customers who have purchased this product may leave a review.