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  5. GAPDH Antibody (HRP) (YA874)

GAPDH Antibody (HRP) (YA874)

Cat. No.: HY-P80954A
COA
SDS
User Guide for Antibodies Technical Support

GAPDH Antibody (HRP) (YA874) is a HRP-conjugated, mouse-derived, anti-GAPDH (HRP) (YA874) IgG1 monoclonal antibody.

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20 μL 해외재고보유
50 μL 해외재고보유
100 μL 해외재고보유
250 μL   견적 받기  
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Top Publications Citing Use of Products
  • WB: Western Blot;
  • IHC-P: Immunohistochemistry-Paraffin;
  • IHC-F: Immunohistochemistry-Frozen;
  • ICC/IF: Immunocytochemistry/Immunofluorescence;
  • IF-Tissue: Immunofluorescence-Tissue;
  • mIHC: Multiplex Immunohistochemical;
  • IP: Immunoprecipitation;
  • ChIP: Chromatin Immunoprecipitation;
  • FC: Flow Cytometry;
  • ELISA: Enzyme Linked Immunosorbent Assay

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  • 제품 설명

제품 설명

GAPDH Antibody (HRP) (YA874) is a HRP-conjugated, mouse-derived, anti-GAPDH (HRP) (YA874) IgG1 monoclonal antibody.
Host

Mouse
Clonality

Monoclonal
분자량
Predicted band size: 36 kDa;
Observed band size: 36 kDa
Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
Species Reactivity
Human, Mouse, Rat
SwissProt ID
Gene ID
Immunogen

A synthetic peptide corresponding to residues near the amino terminus of human GAPDH protein.
Application &
Dilution Ratio
Application Dilution Ratio
WB
WB: Western Blot
1:1000-1:5000
ELISA
ELISA: Enzyme Linked Immunosorbent Assay
1:10000
Sensitivity Endogenous Purity Affinity Purified
Conjugation HRP Modification Unmodified
Isotype IgG1  
Appearance

Solution
Formulation

Supplied in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide, pH 7.3.
선적

Shipping with blue ice.
Verification Image
WB

  • Western blot analysis of extracts from MDA-MB-231 (lane 1) and Raji (lane 2) and Hela (lane 3) and Ramos (lane 4) using beta Tubulin antibody. Proteins were transferred to a PVDF membrane and blocked with 5% nonfat powdered milk in PBST for 2 hour at room temperature. The primary antibody (1/3000) and Loading control antibody (GAPDH, HY-P80954A, 1/3000) was diluted with 5% nonfat powdered milk in PBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/8,000) was incubated for 45min at room temperature.

  • Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), K562 (lane 2, 15 μg), 3T3 (lane 3, 15 μg), C6 (lane 4, 15 μg), and RAW264.7 (lane 5, 15 μg), using GAPDH Mouse mAb (HRP).Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST at 4°C overnight.The primary antibody (1:10000 dilution) and the loading control antibody (beta-Actin, HY-P83730, 1:20000 dilution) were incubated in 5% non-fat milk in TBST for 1 hour at 37°C.Goat Anti-Rabbit IgG-HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

  • HeLa cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with human IFN-α (HY-P70241) at concentration of 20 nM for 30 mins (Serum free). Phospho-IκB-α (Ser32) protein expression was subsequently analyzed by Western blot. The results demonstrated that IFN-α induced phosphorylation of IκB-α (Ser32). Primary antibodies used: Phospho-IκB-α (Ser32) Antibody (HY-P87716) and GAPDH Antibody (HY-P80954A).

  • HeLa cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with human IFN-α (HY-P70241) at concentration of 20 and 50 nM for 30 mins (Serum free). Phospho-STAT3 (Tyr705) protein expression was subsequently analyzed by Western blot. The results demonstrated that IFN-α induced phosphorylation of STAT3 (Tyr705). Primary antibodies used: Phospho-STAT3 (Tyr705) Antibody (HY-P87666) and GAPDH Antibody (HY-P80954A).

  • HeLa cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with Etoposide (HY-13629) at concentration of 30 μM for 2 h and 4 h, respectively. Phospho-ATM (Ser1981) protein expression was subsequently analyzed by Western blot. The results demonstrated that Etoposide induced DNA damage in HeLa cells, indicated by phosphorylation of ATM (Ser1981). Primary antibodies used: Phospho-ATM (Ser1981) Antibody (HY-P87805) and GAPDH Antibody (HY-P80954A).

  • HeLa cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with Calyculin A (HY-18983) at concentration of 100 nM for 30 mins. Phospho-Rb (Thr356) protein expression was subsequently analyzed by Western blot. The results demonstrated that Calyculin A induced phosphorylation of Rb (Thr356). Primary antibodies used: Phospho-Rb (Thr356) Antibody (HY-P81301) and GAPDH Antibody (HY-P80954A).

  • HeLa cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with Calyculin A (HY-18983) at concentration of 10 and 25 nM for 30 mins. Phospho-GSK3 beta (Ser9) protein expression was subsequently analyzed by Western blot. The results demonstrated that Calyculin A induced phosphorylation of GSK3 beta (Ser9). Primary antibodies used: Phospho-GSK3 beta(Ser9) Antibody (HY-P86595) and GAPDH Antibody (HY-P80954A).

  • HeLa cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with Calyculin A (HY-18983) at concentration of 10 nM for 30 mins. Phospho-FAK (Tyr397) protein expression was subsequently analyzed by Western blot. The results demonstrated that Calyculin A induced phosphorylation of FAK (Tyr397). Primary antibodies used: Phospho-FAK (Tyr397) Antibody (HY-P87691) and GAPDH Antibody (HY-P80954A).

  • RAW264.7 cells were cultured in 6 cm dishes to approximately 70% confluence and then treated with LPS (HY-D1056) at concentrations ranging from 0 to 1 μg/mL for 4 h. IRG1 protein expression was subsequently analyzed by Western blot. The results demonstrated that LPS induced IRG1 expression in a dose-dependent manner, confirming the responsiveness of the model. Primary antibodies used: IRG1 Antibody (HY-P84946) and GAPDH Antibody (HY-P80954A).

Background
Function：Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively (PubMed:11724794, PubMed:3170585). 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:11724794, PubMed:3170585). 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)
Subcellular Localization：Cytoplasm, cytosol; Nucleus; Cytoplasm, perinuclear region; Membrane; Cytoplasm, cytoskeleton
Isoforms & Post-Translational Modification：P04406 has 2 isomers: P04406-1: 36053 Da (predicted); P04406-2: 31548 Da (predicted).
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)
Subunit：Homotetramer (PubMed:16239728, PubMed:16510976).
RRID
Database

Entrez Gene: 2597 Human ; 14433 Mouse ; 24383 Rat

SwissProt: P04406 Human ; P16858 Mouse ; P04797 Rat

OMIM: 138400 Human

Research Field

Neuroscience
Synonyms
GAPD, CDABP0047, OK/SW-cl.12, GAPDH, Glyceraldehyde-3-phosphate dehydrogenase, Peptidyl-cysteine S-nitrosylase GAPDH
각종 서류
SDS (251 KB)

COA (205 KB)

User Guide for Antibodies (1077 KB)

GAPDH Antibody (HRP) (YA874) Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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GAPDH Antibody (HRP) (YA874)
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HY-P80954A
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