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Product Info Summary
| SKU: | M00227-2 |
|---|---|
| Size: | 50 µl |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IHC-P, WB |
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Product info
Product Name
Anti-GAPDH Antibody
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.
Reactive Species
M00227-2 is reactive to GAPDH in Human, Mouse, Rat
Calculated molecular weight
36.1 kDa
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.
Application & Images
Applications
M00227-2 is guaranteed for Flow Cytometry, IHC-P, WB Boster Guarantee
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.
Specific Publications For Anti-GAPDH Antibody (M00227-2)
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