Rat Synaptotagmin-1 Antibody

Synaptotagmin‑1 in Rat Neurons. Synaptotagmin-1 was detected in immersion fixed rat hippocampal neurons (E18) using Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to synapses. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Synaptotagmin in Rat Spinal Cord. Synaptotagmin was detected in perfusion fixed frozen sections of rat spinal cord using 8 µg/mL Mouse Anti-Rat Synaptotagmin Monoclonal Antibody (Catalog # MAB4364) overnight at 4 °C. Tissue was stained (red) and counter-stained (green). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Synaptotagmin-1 in SH-SY5Y Human Cell Line. SH-SY5Y human neuroblastoma cells were cultured overnight in the presence of 1 mM Retinoic Acid (Catalog # 0695/50) prior to immersion fixation. Synaptotagmin-1 was detected using a Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364). The cells were stained with the NorthernLights 557-conjugated Donkey Anti-Mouse IgG Affinity-purified Secondary Antibody (red; Catalog # NL007). The cell nuclei were counterstained with DAPI (blue). Synaptotagmin-1 immunoreactivity was localized to synaptic vesicles in the cytoplasm and on the plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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).