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GAPDH Antibody

Purified Mouse Monoclonal Antibody (Mab)

     
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  • 3 - GAPDH Antibody AM8539b
    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell line) cells labeling GAPDH with AM8539b at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG (NA166821) secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm and nucleus staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
  • 14 - GAPDH Antibody AM8539b
    AM8539b staining GAPDH in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
  • 14 - GAPDH Antibody AM8539b
    AM8539b staining GAPDH in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
  • 14 - GAPDH Antibody AM8539b
    AM8539b staining GAPDH in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
  • 14 - GAPDH Antibody AM8539b
    AM8539b staining GAPDH in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
  • 4 - GAPDH Antibody AM8539b
    Overlay histogram showing Hela cells stained with AM8539b(green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AM8539b, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(OJ192088) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was mouse IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
  • 1 - GAPDH Antibody AM8539b
    All lanes : Anti-GAPDH Antibody at 1:8000 dilution Lane 1: C6 whole cell lysate Lane 2: mouse brain lysate Lane 3: NIH/3T3 whole cell lysate Lane 4: Hela whole cell lysate Lane 5: Jurkat whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 36 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
  • 1 - GAPDH Antibody AM8539b
    All lanes : Anti-GAPDH Antibody at 1:8000 dilution Lane 1: Hela whole cell lysate Lane 2: Jurkat whole cell lysate Lane 3: A549 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 36 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
  • 1 - GAPDH Antibody AM8539b
    All lanes : Anti-GAPDH Antibody at 1:8000 dilution Lane 1: Hela whole cell lysate Lane 2: Jurkat whole cell lysate Lane 3: A549 whole cell lysate Lane 4: C6 whole cell lysate Lane 5: NIH/3T3 whole cell lysate Lane 6: mouse brain lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 36 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
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Product info
Application
  • Applications Legend:
  • E=ELISA
  • WB=Western Blotting
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin)
  • IP=Immunoprecipitation
  • IF=Immunofluorescence
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • FC=Flow Cytometry
  • DB=Dot Blot
IF, IHC-P, FC, WB, E
Primary Accession P04406
Reactivity Human, Mouse, Rat
Host Mouse
Clonality monoclonal
Isotype IgG1,k
Clone Names 1653CT401.3.33
Calculated MW 36053 Da
Additional info
Gene ID 2597
Other Names Glyceraldehyde-3-phosphate dehydrogenase, GAPDH, 1.2.1.12, Peptidyl-cysteine S-nitrosylase GAPDH, 2.6.99.-, GAPDH, GAPD
Target/Specificity This GAPDH antibody is generated from a mouse immunized with a recombinant protein between 43-335 amino acids from human GAPDH.
Dilution IF~~1:25
IHC-P~~1:25
FC~~1:25
WB~~1:8000
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
PrecautionsGAPDH Antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name GAPDH
Synonyms GAPD
Function Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). 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. Component of the GAIT (gamma interferon- activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. 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.
Cellular 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.
Research Areas

BACKGROUND

Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). 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. Component of the GAIT (gamma interferon- activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. 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.

REFERENCES

Hanauer A.,et al.EMBO J. 3:2627-2633(1984).
Arcari P.,et al.Nucleic Acids Res. 12:9179-9189(1984).
Tso J.Y.,et al.Nucleic Acids Res. 13:2485-2502(1985).
Tokunaga K.,et al.Cancer Res. 47:5616-5619(1987).
Allen R.W.,et al.J. Biol. Chem. 262:649-653(1987).

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