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  4. GAPDH Antibody (YA3641)

GAPDH Antibody (YA3641) is a mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to GAPDH. It can be used as a loading control antibody.

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Top Publications Citing Use of Products

1 Publications Citing Use of MCE GAPDH Antibody (YA3641)

  • 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
  • Product Detail

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  • Documentation

Description

GAPDH Antibody (YA3641) is a mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to GAPDH. It can be used as a loading control antibody.

Host

Mouse

Clonality

Monoclonal

Molecular Weight
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, Rabbit, Monkey, Pig
SwissProt ID
Gene ID
Immunogen

Purified recombinant fragment of human GAPDH aa 183-333.

Application &
Dilution Ratio
Application Dilution Ratio
WB
WB: Western Blot
1:5000- 1:20000
IHC-P
IHC-P: Immunohistochemistry-Paraffin
1:200-1:1000
ICC/IF
ICC/IF: Immunocytochemistry/Immunofluorescence
1:200-1:1000
FC
FC: Flow Cytometry
1:200-1:400
ELISA
ELISA: Enzyme Linked Immunosorbent Assay
1:10000
Purity affinity purified. Conjugation Non-conjugated
Modification Unmodified Isotype IgG1
Appearance

Liquid

Formulation

Supplied in PBS with 0.05% sodium azide.

Storage & Stability

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping

Shipping with blue ice.

Verification Image
ALL WB IHC-P FC ICC
  • Western blot analysis of extracts from Jurkat (lane 2(20μg), Hela (lane 3(20μg), HepG2 (lane 4(20μg), THP-1 (lane 5(20μg) using GAPDH Antibody(HY-P83944). Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/5000) and Loading control antibody (Beta Actin, HY-P80993, 1/10,000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (HY-P8004 ,1/10,000) was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue using GAPDH Antibody (HY-P83944, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue using GAPDH Antibody (HY-P83944, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using GAPDH Antibody (HY-P83944, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human stomach tissue using GAPDH Antibody (HY-P83944, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using GAPDH Antibody (HY-P83944, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue using GAPDH Antibody (HY-P83944, 1/500). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of 1X106 HeLa cells labeling GAPDH Antibody (HY-P83944, red). Cells were fixed with 4% paraformaldehyde. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. AF 488-conjugated AffiniPure Goat Anti- Mouse IgG H&L (HY-P8005) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Mouse IgG Isotype Control (HY-P80757, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

  • Immunocytochemistry analysis of HeLa cells labeling GAPDH with GAPDH antibody (HY-P83944) at 1/300 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with GAPDH antibody (HY-P83944) at 1/300 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Mouse IgG H&L(HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Immunocytochemistry analysis of HeLa cells labeling GAPDH with GAPDH antibody (HY-P83944) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with GAPDH antibody (HY-P83944) at 1/500 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Mouse IgG H&L(HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

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
Synonyms
G3PD; GAPD; MGC88685; Glyceraldehyde-3-phosphate dehydrogenase
Documentation
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 (YA3641)
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HY-P83944
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