Human/Mouse/Rat VAMP-2 Antibody

Detection of Human, Mouse, and Rat VAMP‑2 by Western Blot.

Western blot shows lysates of Nalm-6 human Pre-B acute lymphocytic leukemia cell line, SH-SY5Y human neuroblastoma cell line, mouse brain tissue, and rat testis tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat VAMP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5136) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for VAMP-2 at approximately 13 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

VAMP-2 in Rat Spinal Cord.

Vesicle-Associate Membrane Protein 2 (VAMP-2) was detected in perfusion-fixed frozen sections of rat spinal cord using a Goat Anti-Human/Mouse/Rat VAMP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5136). The cells were stained with the NorthernLights 557-conjugated Donkey Anti-Goat IgG Affinity-purified Secondary Antibody (red; Catalog # NL001). The cell nuclei were counterstained with DAPI (blue). VAMP-2 immunoreactivity was localized to synaptic vesicles. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of VAMP-2 by Western Blot

Stx2 deletion enhances exocytosis by facilitating trans-SNARE complex assembly and cis-SNARE complex disassembly.a Stx2 deletion promotes the formation of profusion trans-SNARE complexes. SNARE complexes were co-immunoprecipitated (co-IP) from human islets lysates by antibody against Stx1a and Stx3. Native primary antibody-specific secondary antibodies were used to eliminate interference by the heavy- and light-chain IgG fragments of initial IP assay. Representative blots are shown in a and quantification shown in Supplementary Fig. 3a (n = 4 independent experiments). b Stx2 inhibits the transition of early assembly intermediates to fusogenic SNARE complexes by substituting for the fusogenic syntaxins (Stx1a, Stx3) as detected by native-PAGE. Samples were loaded with 5× native loading buffer into 12% non-denaturing polyacrylamide gel without SDS. This is because Stx2 directly bound not only cognate SNAREs but also the priming factors, Munc18a and Munc13 (in Supplementary Fig. 3b) (n = 3 independent experiments). c, d Stx2 deletion enhances disassembly of cis-SNARE complexes. Native SNARE complexes isolated from INS832/13 by antibody against VAMP2 or VAMP8 were disassembled by recombinant NSF and alpha-SNAP in the absence or presence of Stx2. 10 mM EDTA was used to inactivate ATP hydrolysis by chelating Mg2+. SDS-resistant complexes disassemble completely when the sample was heated to 100 °C before electrophoresis. SNARE complexes were visualized by SDS/PAGE and immunoblotting using antibody against SNAP25. Representative blots are shown in c and quantification in d. Values are mean ± s.d. from 4 independent experiments (two-tailed unpaired Student’s t-test). Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36316316), licensed under a CC-BY license. Not internally tested by R&D Systems.