Anti-GAPDH Rabbit Monoclonal Antibody


SKU M00227
Size 100ug/vial
Reactivity Human, Mouse, Rat
Clonality Monoclonal
Clone Number BH-7
Host Rabbit
Applications Flow Cytometry, IP, IF, IHC, ICC, WB

Overview

Product Name Anti-GAPDH Rabbit Monoclonal Antibody
SKU/Catalog Number M00227
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.
Size 100ug/vial
Description Anti-GAPDH Rabbit Monoclonal Antibody tested for Flow Cytometry, IP, IF, IHC, ICC, WB in Human, Mouse, Rat
Cite This Product Anti-GAPDH Rabbit Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00227)
Host Rabbit
Contents/Buffer Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Form Liquid
Immunogen A synthesized peptide derived from human GAPDH
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: 0.5-1mg/ml, actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
WB 1:3000-1:10000
IHC 1:100-1:500
ICC/IF 1:100-1:250
IP 1:50
Flow Cytometry 1:50

Images And Assay Conditions

Immunofluorescent analysis of Hela cells, using GAPDH Antibody

Immunohistochemical analysis of paraffin-embedded (1) Rat colon; (2) Mouse testis; (3) Human uterus; (4) Human kidney, using GAPDH Antibody (M00227)
GAPDH was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-GAPDH Antibody (M00227)overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Western blot analysis of GAPDH expression in (1) Hela cell lysate; (2)Jurkat cell lysate; (3)Mouse kidney lysate; (4) Mouse spleen lysate; (5) RAW 2647 cell lysate; (6) Rat brain lysate with GAPDH Antibody (M00227).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH monoclonal antibody (Catalog # M00227) overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAPDH

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P04406
Gene Name GAPDH
Protein Name Glyceraldehyde-3-phosphate dehydrogenase
Alternative Names Glyceraldehyde-3-phosphate dehydrogenase;GAPDH;1.2.1.12;Peptidyl-cysteine S-nitrosylase GAPDH;2.6.99.-;GAPDH;GAPD;CDABP0047, OK/SW-cl.12;
Subcellular Localization Cytoplasm, cytosol . Nucleus . Cytoplasm, perinuclear region . Membrane . Cytoplasm, cytoskeleton . Translocates to the nucleus following S- nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions. .
Molecular Weight 36053 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein 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. .
Research Areas Human, Mouse, Rat

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Monoclonal antibody for GAPDH detection. Host: Rabbit.Size: 100ug/vial. Tested applications: Flow Cytometry, IP, IF, IHC, ICC, WB. Reactive species: Human, Mouse, Rat GAPDH information: Molecular Weight: 36053 MW; Subcellular Localization: Cytoplasm, cytosol . Nucleus . Cytoplasm, perinuclear region . Membrane . Cytoplasm, cytoskeleton . Translocates to the nucleus following S- nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
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In stock
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M00227
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$299.00

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