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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human VAMP8 (N terminal). The exact sequence is proprietary.
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Alternative versions available:
Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.
Our Abpromise guarantee covers the use of ab76021 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000 - 1/20000. Detects a band of approximately 17 kDa (predicted molecular weight: 11 kDa).|
|IP||1/20 - 1/50.|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).
|Flow Cyt||1/80 - 1/150.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/100 - 1/250.|
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling VAMP8 with purified ab76021 at a dilution of 1/250. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/Immunofluorescence analysis of PC-12 cells labelling VAMP8 with purified ab76021 at a dilution of 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Flow Cytometry analysis of HeLa cells labelling VAMP8 with purified ab76021 at a dilution of 1/150 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
ab76021 (purified) at 1/50 immunoprecipitating VAMP8 in HEK293 whole cell lysate.
Lane 1 (input): HEK293 whole cell lysate (10µg)
Lane 2 (+): ab76021 + HEK293 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76021 in HEK293 whole cell lysate.
For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labelling VAMP8 with unpurified ab76021 at a dilution of 1/100. A HRP/AP polymerized secondary antibody was used.
Immunohistochemical analysis of Human lacrimal gland tissue staining VAMP8 with unpurified ab76021.
Antigen retrieval was performed using antigen retrieval solution in a microwave. Sections were blocked with 10 goat serum for 30 minutes and incubated with primary antibody (1/100) overnight at 4°C. Staining was detected using DAB.