Human/Rat Syntaxin 5 Antibody

  
  • Species Reactivity
    Human, Rat
  • Specificity
    Detects human Syntaxin 5 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross‑reactivity with recombinant human (rh) Syntaxin 7, rhSyntaxin 8, and rhSytaxin 12 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Syntaxin 5 isoform 2
    Met55-Thr300
    Accession # Q13190
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    25 µg/mL
    See below
  • Immunohistochemistry
    3-15 µg/mL
    See below
  • Immunocytochemistry
    1-15 µg/mL
    See below
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.
Data Examples
Detection of Human Syntaxin 5 by Western Blot. Western blot shows lysates of T47D human breast cancer cell line and Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Rat Syntaxin 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5687) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Syntaxin 5 at approximately 35 and 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Syntaxin 5 in SH-SY5Y Human Cell Line. Syntaxin 5 was detected in immersion fixed SH-SY5Y human neuroblastoma cell line using Goat Anti-Human/Rat Syntaxin 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5687) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
Syntaxin 5 in Human Brain. Syntaxin 5 was detected in immersion fixed paraffin-embedded sections of human brain (substantia nigra) using Goat Anti-Human/Rat Syntaxin 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5687) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
Syntaxin 5 in Rat Brain. Syntaxin 5 was detected in immersion fixed frozen sections of rat brain using Goat Anti-Human/Rat Syntaxin 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5687) at 3 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to neuronal cytoplasm. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Human Syntaxin 5 by Simple WesternTM. Simple Western lane view shows lysates of T47D human breast cancer cell line and Raji human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. Specific bands were detected for Syntaxin 5 at approximately 39 and 47 kDa (as indicated) using 25 µg/mL of Goat Anti-Human/Rat Syntaxin 5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5687) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
Background: Syntaxin 5
Syntaxin 5 (STX5) is a predicted 41‑42 kDa member of the syntaxin family of proteins. It is a t‑SNARE that is widely expressed (but not in striated muscle), and embedded in either the cis‑Golgi network (a 37 kDa form) or the ER (the 46 kDa form). STX5 is involved in the regeneration and maintenance of the Golgi apparatus after mitosis. Human STX5 is a type IV single‑pass transmembrane protein (very long cytoplasmic N‑terminus) that is 355 amino acids (aa) in length. It contains an ER retrieval signal (Arg4Lys5Arg6), a t‑SNARE domain (aa 263‑325) with a coiled‑coil region (aa 287‑318), and a C‑terminal transmembrane sequence. The 35 kDa short form exhibits an alternate start site at Met55. Two other potential splice variants show either the Met55, or a Met97 start site coupled to a 19 aa substitution for aa 303‑355. Over aa 55‑300, human STX5 shares 94% aa identity with mouse STX5.
  • Entrez Gene IDs:
    6811 (Human); 56389 (Mouse); 65134 (Rat)
  • Alternate Names:
    SED5; STX5; STX5ASED5; Syntaxin 5; syntaxin 5A; syntaxin-5
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