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Affinity purified polyclonal antibody
- SPECIFICATION
- CITATIONS
- PROTOCOLS
- BACKGROUND
Application 
| WB |
|---|---|
| Primary Accession | P63045 |
| Reactivity | Mouse, Rat |
| Host | Rabbit |
| Clonality | Polyclonal |
| Calculated MW | 12691 Da |
| Homology | Human, bovine, Macaca mulata - 25/26 amino acid residues identical. |
| Gene ID | 24803 |
|---|---|
| Other Names | Vesicle-associated membrane protein 2, VAMP-2, Synaptobrevin-2, Vamp2, Syb2 |
| Related products for control experiments | Control peptide antigen (supplied with the antibody free of charge). |
| Target/Specificity | GST fusion protein with sequence SATAATVPPAAPAGEGGPPAPPPNLT, corresponding to amino acid residues 2-27 of rat VAMP-2 (Accession P63045).ֲ ֲ Cytoplasmic, N-terminus. |
| Dilution | WB~~1:200-1:2000 |
| Peptide Confirmation | Confirmed by DNA sequence and SDS-PAGE. |
| Format | Affinity purified antibody, lyophilized powder |
| Reconstitution | 50 µl or 0.2 ml deionized water, depending on the sample size. |
| Antibody Concentration After Reconstitution | 1 mg/ml. |
| Buffer After Reconstitution | Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 5% sucrose, 0.025% NaN3. |
| Storage Before Reconstitution | Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C. |
| Storage After Reconstitution | The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min). |
| Control Antigen Storage Before Reconstitution | Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C. |
| Control Antigen Reconstitution | 100 µl PBS. |
| Control Antigen Storage After Reconstitution | -20ºC. |
| Preadsorption Control | 3 µg fusion protein per 1 µg antibody. |
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Provided below are standard protocols that you may find useful for product applications.
Background
VAMP-2 (also known as synaptobrevin-2) is a member of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein superfamily. The family includes 36 members in humans and is characterized by the SNARE motif, an evolutionarily conserved stretch of 60–70 amino acids that are arranged in heptad repeats1,2. SNARE proteins are involved in exocytosis and intracellular vesicle trafficking and are essential for cell growth, hormone secretion and neurotransmission, processes that require rapid, targeted, and regulated membrane fusion1,2. SNAREs can be roughly divided into vesicular (v-SNAREs) and target (t-SNAREs) based on their distribution on the transport vesicle or target membrane respectively. Thus, assembly of cognate v-/t-SNAREs between two opposing membranes generates trans-SNARE complexes, which bring the lipid bilayers in close proximity and drive membrane fusion. VAMP-2, like most SNAREs, is a type IV membrane protein with a relatively large N-terminus containing the SNARE motif located in the cytoplasmic side and a transmembrane domain located close to the C-terminus that functions as an anchor1,2. VAMP-2 has been extensively studied for its role on neuronal and neuroendocrine cell exocytosis where it functions as the vesicle membrane protein v-SNARE, which together with the plama membrane t-SNARE protein Syntaxin 1 and the membrane-associated SNAP-25 (synaptosome-associated protein 25 kDa), forms a trimeric, four-helical complex, which drives fusion of the two opposing bilayers1,2. VAMP-2 is the target of the tetanus neurotoxin (TeNT) and of several botulinum neurotoxin (BoNT) types: type B, D, F, and G. The neurotoxins cause specific proteolytic degradation of the VAMP-2 protein, which in turn causes SNARE complex disruption and inhibition of neurotransmitter release3. Abgent is pleased to offer a highly specific antibody directed against the intracellular N-terminus domain of rat VAMP-2. Anti-VAMP-2 antibody (#AG1191) can be used in western blot and immunohistochemical applications, and was designed to recognize VAMP-2 from rat, mouse and human samples.
References
References 1. Jahn, R. and Scheller, R.H. (2006) Nat. Rev. Mol. Cell Biol. 7, 631. 2. Südhof, T.C. and Rothman, J.E. (2009) Science 323, 474. 3. Schiavo, G. et al. (2000) Physiol. Rev. 80, 717.
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