The epitope for this antibody has been localized to the amino terminus (residues 1-25) of Syntaxin 6.
Mouse Brain Tissue Extract, rat Brain Tissue Extract.
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab140607 as a replacement.
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Immunocytochemistry/ Immunofluorescence - Syntaxin 6 antibody [3D10] (ab12370)This image is courtesy of an anonymous Abreview
ab12370 at 1/200 staining a mouse macrophage cell line by ICC/IF. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 24 hours. A FITC conjugated donkey anti-mouse antibody was used as the secondary.
Syntaxin 6 was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 10µg of Mouse monoclonal to Syntaxin 6 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab12370. Secondary: Protein G-HRP at 1/500 dilution. Band: 25kDa:Syntaxin 6.
Overlay histogram showing PC12 cells stained with ab12370 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab12370, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.