Human Syntaxin 4 Antibody Summary
Accession # Q12846
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Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Syntaxin 4 by Western Blot. Western blot shows lysates of A172 human glioblastoma cell line and SH-SY5Y human neuroblastoma cell line. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Syntaxin 4 Monoclonal Antibody (Catalog # MAB7894) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Syntaxin 4 at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Syntaxin 4 in U‑87 MG Human Cell Line. Syntaxin 4 was detected in immersion fixed U-87 MG human glioblastoma/astrocytoma cell line using Mouse Anti-Human Syntaxin 4 Monoclonal Antibody (Catalog # MAB7894) at 25 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 plasma membranes and focal contacts. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Syntaxin 4 by Simple WesternTM. Simple Western lane view shows lysates of SH‑SY5Y human neuroblastoma cell line, loaded at 0.5 mg/mL. A specific band was detected for Syntaxin 4 at approximately 42 kDa (as indicated) using 2.5 µg/mL of Mouse Anti-Human Syntaxin 4 Monoclonal Antibody (Catalog # MAB7894). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Human Syntaxin 4 Specificity Using Knockout Cell Line. Western blot shows lysates of HAP1 human near-haploid cell line and Syntaxin 4 knockout HAP1 cell line (KO). Nitrocellulose membrane was probed with 0.25 µg/mL of Mouse Anti-Human Syntaxin 4 Monoclonal Antibody (Catalog # MAB7894) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody. A specific band was detected for Syntaxin 4 at approximately 34 kDa (as indicated) in the parental HAP1 cell line, but is not detectable in knockout HAP1 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-4 by Immunoprecipitation. Immunoprecipitation was performed on cell lysate of HAP1 human near-haploid cell line using 2.0 μg of Mouse Anti-Human Syntaxin-4 Monoclonal Antibody (Catalog # MAB7894) pre-coupled to protein G or protein A beads. Immunoprecipitated Syntaxin-4 was detected with an Anti-Human Syntaxin-4 Monoclonal Antibody (Catalog # MAB7894) The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Syntaxin 4
Syntaxin 4 (STX4) is a widely expressed type IV transmembrane protein that is a member of the syntaxin family of t‑SNARE (target‑Soluble NSF [N‑ethylmaleimide‑sensitive factor] Attachment REceptor) membrane fusion proteins (1, 2). For membrane fusion, VAMPs (Vesicle‑Associated Membrane Proteins, also called Synaptotagmins) as v‑SNAREs on donor (vesicle) membranes interact with t‑SNAREs and SNAPs (Soluble NSF Attachment Proteins) on target (cell surface) membranes, with SM (Sec1/Munc18-like) accessory proteins acting as catalysts for fusion (2). Human Syntaxin 4 cDNA encodes 297 amino acids (aa) including basolateral targeting motif for epithelial cells (aa 24‑29), a coiled‑coil domain termed Habc (aa 43‑163), an S‑nitrosylation site which regulates fusion (aa 141), a hinge region, a t‑SNARE domain (aa 200‑262), and a C‑terminal transmembrane domain (aa 276‑296) (1‑4). Within aa 1‑183, human Syntaxin 4 shares 89% aa sequence homology with mouse and rat, and 94‑95% with canine, bovine, equine, porcine, feline and ovine Syntaxin 4. Among its known roles, Syntaxin 4 and SNAP‑23 or SNAP‑25 associate with VAMP‑2 for fusion of GLUT4‑containing vesicles with the plasma membrane in muscle and adipose cells (1, 5). It is required for exocytosis of insulin from pancreatic beta ‑cells (3, 6, 7). With SNAP23 and VAMP‑3, it allows endosome recycling to extend podosomes during macrophage migration (7‑9). It establishes polarization of epithelial cells, specifying basolateral sorting of proteins (4, 10). It mediates postsynaptic activity‑dependent exocytosis in neural cell dendritic spines (11). It is involved in exocytosis of ANP from cardiomyocytes (12). In FAS‑mediated apoptosis, it allows translocation of acid sphingomyelinase to the cell surface (13). Syntaxin 4 binds F‑actin and gelsolin, linking the plasma membrane with, or releasing it from, the actin cytoskeleton (6, 7).
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