Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human

Applications

Knockout Validated, Western Blot, Immunocytochemistry, Simple Western, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 884637
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Product Specifications

Immunogen

E. coli-derived recombinant human Syntaxin 4
Met1-Glu183
Accession # Q12846

Specificity

Detects human Syntaxin 4 in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Syntaxin 4 Antibody

Detection of Human Syntaxin 4 antibody by Western Blot.

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 antibody in U-87 MG Human Cell Line by Immunocytochemistry (ICC).

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 antibody by Simple WesternTM.

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).

Applications for Human Syntaxin 4 Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed U‑87 MG human glioblastoma/astrocytoma cell line

Immunoprecipitation

2 µg/500 µg cell lysate
Sample: Cell lysate of HAP1 human near-haploid cell line.

Knockout Validated

Syntaxin 4 is specifically detected in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line.

Simple Western

2.5 µg/mL
Sample: SH‑SY5Y human neuroblastoma cell line

Western Blot

0.25 µg/mL
Sample: A172 human glioblastoma cell line and SH‑SY5Y human neuroblastoma cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References

  1. Jagadish, M.N. et al. (1996) Biochem. J. 317:945.
  2. Sudhof, T.C. and J.E. Rothman 2009 Science 323:474.
  3. Wiseman, D.A. et al. (2011) J. Biol. Chem. 286:16344.
  4. Reales, E. et al. (2011) PLoS One 6:e21181.
  5. Vicogne, J. et al. (2006) Proc. Natl. Acad. Sci. USA 103:14761.
  6. Jewell, J.L. et al. (2008) J. Biol. Chem. 283:10716.
  7. Kalwat, M.A. et al. (2012) Mol. Endocrinol. 26:128.
  8. Veale, K.J. et al. (2010) Commun. Integr. Biol. 4:44.
  9. Veale, K.J. et al. (2010) Exp. Cell Res. 317:1817.
  10. Low, S.H. et al. (2006) Mol. Biol. Cell 17:977.
  11. Kennedy, M.J. et al. (2010) Cell 141:524.
  12. Ferlito, M. et al. (2010) J. Mol. Cardiol. 49:791.
  13. Perotta, C. et al. (2010) J. Biol. Chem. 285:40240.

Alternate Names

NY-REN-31, p35-2, STX4

Entrez Gene IDs

6810 (Human); 20909 (Mouse); 81803 (Rat)

Gene Symbol

STX4

UniProt

Additional Syntaxin 4 Products

Product Documents for Human Syntaxin 4 Antibody

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Product Specific Notices for Human Syntaxin 4 Antibody

For research use only

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Protocols

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