ICC/IF image mouse anti GAPDH monoclonal antibody 1D4 5157

mouse anti-GAPDH monoclonal antibody (1D4) 5157

$100.00 – $2,600.00

Antibody summary

Mouse monoclonal to GAPDH
Suitable for: WB, ICC/IF, IHC
Reacts with: human, mouse, rat
Isotype: IgM
100 µg, 25 µg, 1 mg

SKU: 5157parent Categories: antibody, loading control Tag: GAPDH

Weight	1 lbs
Dimensions	9 × 5 × 2 in
host	mouse
isotype	IgM
clonality	monoclonal
concentration	1 mg/mL
applications	ICC/IF, IHC, WB
reactivity	human, mouse, rat
available sizes	1 mg, 100 µg, 25 µg

mouse anti-GAPDH monoclonal antibody (1D4) 5157

antibody
Database link: human P04406 mouse P16858 rat P04797
Tested applications WB,IHC,IHC,ICC/IF
Recommended dilutions WB: 1:1000 IF/IHC: 1:100
Immunogen Full length protein purified from pig blood cells
Size and concentration 25, 100, 1000µg and 1 mg/mL
Form liquid
Storage Instructions 2-8°C for short term, for longer term at -20°C. Avoid freeze / thaw cycles.
Storage buffer PBS, 50% glycerol, 0.04% NaN₃
Purity affinity purified
Clonality monoclonal
Isotype IgM
Compatible secondaries goat anti-mouse IgG, H&L chain specific, peroxidase conjugated polyclonal antibody 5486 goat anti-mouse IgG, H&L chain specific, biotin conjugated, Conjugate polyclonal antibody 2685 goat anti-mouse IgG, H&L chain specific, FITC conjugated polyclonal antibody 7854 goat anti-mouse IgG, H&L chain specific, peroxidase conjugated polyclonal antibody, crossabsorbed 1706 goat anti-mouse IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1716 goat anti-mouse IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1721
Isotype control Mouse monocolonal IgM- 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

Immunofluorescent analysis of HeLa cells stained with mouse mAb to GAPDH, 5157, dilution 1:100 in green. Blue is Hoechst staining of nuclear DNA. The 5157 antibody produces strong cytoplasmic staining of healthy cells.

Chromogenic immunostaining of a NBF fixed paraffin embedded human pancreas section with mouse mAb to GAPDH, 5157, dilution 1:1,000, detected with DAB (brown) using the Vector labs Elite 8718C-HRP detection and goat anti-mouse IgM secondary with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. The GAPDH antibody strongly labels pancreatic exocrine glandular cells. This antibody performs well in testing with both 4% PFA and standard NBF fixed tissues.

Western blot analysis of cell line lysates probed with mouse mAb to GAPDH, 5157, 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 blot loading control.

Publications

pmid	title	authors	citation
37961202	LOXL2 inhibition ameliorates pulmonary artery remodeling in pulmonary hypertension.	Jochen Steppan, Huilei Wang, Kavitha Nandakumar, Alan Poe, Lydia Pak, Travis Brady, Mahin Gadkari, Dan E Berkowitz, Larissa A Shimoda, Lakshmi Santhanam	bioRxiv N/A:N/A
37918959	E4F1 and ZNF148 are transcriptional activators of the -57A > C and wild-type TERT promoter.	Boon Haow Chua, Nurkaiyisah Zaal Anuar, Laure Ferry, Cecilia Domrane, Anna Wittek, Vineeth T Mukundan, Sudhakar Jha, Falk Butter, Daniel G Tenen, Pierre-Antoine Defossez, Dennis Kappei	Genome Res 33:1893-1905
37769126	Multiomics analyses reveal dynamic bioenergetic pathways and functional remodeling of the heart during intermittent fasting.	Thiruma V Arumugam, Asfa Alli-Shaik, Elisa A Liehn, Sharmelee Selvaraji, Luting Poh, Vismitha Rajeev, Yoonsuk Cho, Yongeun Cho, Jongho Kim, Joonki Kim, Hannah L F Swa, David Tan Zhi Hao, Chutima Rattanasopa, David Yang-Wei Fann, David Castano Mayan, Gavin Yong-Quan Ng, Sang-Ha Baik, Karthik Mallilankaraman, Mathias Gelderblom, Grant R Drummond, Christopher G Sobey, Brian K Kennedy, Roshni R Singaraja, Mark P Mattson, Dong-Gyu Jo, Jayantha Gunaratne	Elife 12:N/A
37748675	Astrocyte lipocalin-2 modestly effects disease severity in a mouse model of multiple sclerosis while reducing mature oligodendrocyte protein and mRNA expression.	Sergey Kalinin, Douglas L Feinstein	Neurosci Lett 815:137497
37716491	Panax notoginseng saponins inhibits oxidative stress- induced human nucleus pulposus cell apoptosis and delays disc degeneration in vivo and in vitro.	Danqing Guo, Miao Yu, Huizhi Guo, Min Zeng, Yang Shao, Wei Deng, Qiuli Qin, Yongxian Li, Shuncong Zhang	J Ethnopharmacol 319:117166

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.