Rat Synaptotagmin-1 Antibody

Western Blot Shows Human Synaptotagmin-1 Specificity Using Knockout Cell Line.

Western blot shows lysates of HCT 116 human colorectal carcinoma cell line and Synaptotagmin-1 knockout HCT 116 cell line (KO). Nitrocellulose membrane was probed with 0.5 µg/mL of Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364) followed by HRP-conjugated donkey anti-sheep IgG Secondary Antibody. A specific band was detected for Synaptotagmin-1 at approximately 63 kDa (as indicated) in the parental HCT 116 cell line, but is not detectable in knockout HCT 116 cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of Synaptotagmin-1 by Immunoprecipitation.

HCT 116 lysates were prepared and immunoprecipitation was performed using 2.0 µg of Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364) pre-coupled to Dynabeads protein G. Immunoprecipitated Synaptotagmin-1 was detected with a Rabbit Anti-Synaptotagmin-1 antibody. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Synaptotagmin-1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

HCT 116 WT and Synaptotagmin-1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364) followed by incubation with a goat anti-mouse Alexa-fluor 555 coupled secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody concentration used: 1 µg/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).