Anti-GAPDH Antibody

Product Name

Anti-GAPDH Antibody

View all GAPDH Antibodies

SKU/Catalog Number

M00227-2

Size

50 µl

Description

Boster Bio Anti-GAPDH Antibody (Catalog # M00227-2). Tested in WB, Flow Cytometry, IHC-P application(s). This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Maintain 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.

Cite This Product

Anti-GAPDH Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00227-2)

Host

Mouse

Contents

Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide.

Clonality

Monoclonal

Clone Number

1653CT614.35.51

Isotype

IgG1,k

Immunogen

This GAPDH antibody is generated from a mouse immunized with a recombinant protein between 43-335 amino acids from human GAPDH.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Reactive Species

M00227-2 is reactive to GAPDH in Human, Mouse, Rat

Applications

M00227-2 is guaranteed for Flow Cytometry, IHC-P, WB Boster Guarantee

Observed Molecular Weight

Calculated molecular weight

36.053kDa

Background of GAPDH

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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

WB: 1:8000
IHC-P: 1:25
FC: 1:25

Validation Images & Assay Conditions

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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.

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M00227-2 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.

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M00227-2 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.

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Overlay histogram showing Hela cells stained with M00227-2 (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 (M00227-2, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed 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.

Gene/Protein Information For GAPDH (Source: Uniprot.org, NCBI)

Alternative Names

1D4 GAPDH; aging-associated gene 9 protein; Cytoplasm Marker; EC 1.2.1; EC 1.2.1.12; EC 2.6.99.-; G3PD; G3PD; G3PDH; GAPD; GAPDH; GAPDPeptidyl-cysteine S-nitrosylase GAPDH; glyceraldehyde 3-phosphate dehydrogenase; glyceraldehyde-3-phosphate dehydrogenase; MGC102544; MGC102546; MGC103190; MGC103191; MGC105239; MGC127711; MGC88685; OCAS, p38; OCT1 coactivator in S phase; Peptidyl-cysteine S-nitrosylase GAPDH GAPDH G3PD, GAPD, HEL-S-162eP glyceraldehyde-3-phosphate dehydrogenase glyceraldehyde-3-phosphate dehydrogenase|OCAS, p38 component|Oct1 coactivator in S phase, 38 Kd component|aging-associated gene 9 protein|epididymis secretory sperm binding protein Li 162eP|peptidyl-cysteine S-nitrosylase GAPDH

For more info on GAPDH, check out the GAPDH Infographic

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In this infographic, you will see the following information for GAPDH: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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