Western blot analysis of extracts from various cell lines using Syntaxin 5 Antibody.
|REACTIVITY||H M R|
|MW (kDa)||35, 42|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
Syntaxin 5 Antibody recognizes endogenous levels of total syntaxin 5 protein. This antibody also cross-reacts with an unidentified protein of 120 kDa.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly165 of human syntaxin 5 protein. Antibodies are purified by protein A and peptide affinity chromatography.
The membrane protein syntaxin 5 (STX5) is a key component of soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor (SNARE) complexes that regulate cellular protein transport, vesicle docking, and membrane fusion (1). Syntaxin 5 protein is found as a 42 kDa ("long") protein localized to the Golgi complex and endoplasmic reticulum, and a “short” 35 kDa isoform localized primarily to the Golgi (2,3). Formation of the syntaxin 5 SNARE complex, which also includes proteins Sec22B, Bet1, GOSR1, GOSR2, and Ykt6, allows for regulation of ER-to-Golgi transport, intra-Golgi transport, and endosome-to-Golgi retrograde transport (4-6). Research studies indicate that the syntaxin 5 SNARE complex also plays an essential role in autophagy following autophagosome formation. Intracellular protein transport mediated by the syntaxin 5 complex is required for transport and localized activity of lysosomal proteases. The experimental reduction or deletion of syntaxin 5 complex components results in non-functional lysosomes and accumulation of autophagosomes (7).
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