Human/Rat Syntaxin 5 Antibody

Catalog #: AF5687 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF5687
AF5687-SP

Detection of Human Syntaxin 5 by Western Blot.

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Human/Rat Syntaxin 5 Antibody Summary

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 either lyophilized or 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.

Scientific Data

Western Blot

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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

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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

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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

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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.

Simple Western

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Detection of Human Syntaxin 5 by Simple Western^TM. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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.

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