Human/Mouse/Rat Syntaxin-BP1 Antibody

Syntaxin-BP1 in Mouse Brain. Syntaxin-BP1 was detected in perfusion fixed frozen sections of mouse brain (hippocampus) using Human/Mouse/Rat Syntaxin-BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5675) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Human/Mouse/Rat Syntaxin-BP1 by Western Blot. Western blot shows lysates of rat brain tissue and spinal cord tissue. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat Syntaxin-BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5675) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Syntaxin-BP1 at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Western Blot Shows Syntaxin‑BP1 Specificity Using Knockout Cell Line. Western blot shows lysates of U‑87 MG human glioblastoma/astrocytoma parental cell line and Syntaxin‑BP1 knockout U-87 MG cell line (KO). Nitrocellulose membrane was probed with 1 ug/mL of Goat Anti-Human/Mouse/Rat Syntaxin‑BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5675) followed by HRP-conjugated secondary antibody. A specific band was detected for Syntaxin‑BP1 at approximately 67 kDa (as indicated) in the parental U-87 MG cell line, but is not detectable in knockout U-87 MG 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 Syntaxin‑BP1 by Immunoprecipitation. U‑87 MG human glioblastoma/astrocytoma cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of Goat Anti-Human/Mouse/Rat Syntaxin‑BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5675) pre-coupled to Dynabeads Protein G. Immunoprecipitated Syntaxin‑BP1 was detected in Western Blot with a Rabbit Syntaxin-BP1 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).

Syntaxin‑BP1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. U‑87 MG human glioblastoma/astrocytoma parental cell line WT and Syntaxin‑BP1 U-87 MG KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with Goat Anti-Human/Mouse/Rat Syntaxin‑BP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5675) followed by incubation with an Alexa-fluor 555 conjugated 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 ug/mL. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).