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VAMP1/2/3 antibody - 104 203 K.D.

Vamp1, 2 and 3 are major vesicle protein involved in fusion
Rabbit polyclonal purified antibody
Cat. No.: 104 203
Amount: 50 µg
Price: $455.00
Cat. No. 104 203 50 µg specific antibody, lyophilized. Affinity purified with the immunogen. Albumin and azide were added for stabilization. For reconstitution add 50 µl H2O to get a 1mg/ml solution in PBS. Then aliquot and store at -20°C to -80°C until use.
Antibodies should be stored at +4°C when still lyophilized. Do not freeze!
Applications
 
WB: 1 : 1000 up to 1 : 5000 (AP staining) (see remarks) gallery  
IP: not recommended
ICC: 1 : 500 (see remarks) gallery  
IHC: 1 : 200 gallery  
IHC-P: 1 : 500 gallery  
DNA-PAINT: yes (see remarks)
ELISA: yes (see remarks)

Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section.

Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section.

Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section.

Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section.

Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.

In antibody-based DNA-PAINT (Point Accumulation in Nanoscale Topography), a short oligomeric docking strand is coupled to a specific antibody. The transient association of the fluorophore to the antibody is mediated by the pairing of a short fluorescently labeled complementary imager DNA strand. DNA-PAINT allows super-resolution imaging and the imaging of a huge number of antibodies on the same biological sample in a single multiplex experiment. For additional experimental details refer to the Remarks section.

Enzyme-linked immunosorbent assay (ELISA); a frequently employed method to quantify target-molecules in solution. The detection of some proteins requires special solubilization steps. For further information, please refer to the „Remarks“ section.

Immunogen Recombinant protein corresponding to AA 1 to 82 from rat Cellubrevin (UniProt Id: P63025)
Reactivity Reacts with: human (P23763, P63027, Q15836), rat (Q63666, P63045, P63025), mouse (Q62442, P63044, P63024), zebrafish.
Other species not tested yet.
Specificity Specific for all three isoforms VAMP 1, 2 and 3. K.D. validated PubMed: 31128202
Remarks

WB: This antibody detects the tetanus and Botulinum B toxin cleavage product in toxin treated tissue homogenates.
ICC: Methanol fixation is recommended. This antibody requires an additional acetone fixation step according to Annaert et al., 1997
This antibody is recommended for ICC of non-neuronal cells.
DNA-PAINT: This antibody has been successfully used for DNA-PAINT application (see Unterauer et al., 2024; PMID: 38552614).
ELISA: Suitable as detector antibody for sandwich-ELISA with cat. no. 104 211 as capture antibodies. The ELISA-protocol for membrane proteins is recommended.

Data sheet 104_203.pdf

References for VAMP1/2/3 - 104 203

The endothelial diapedesis synapse regulates transcellular migration of human T lymphocytes in a CX3CL1- and SNAP23-dependent manner.
Schoppmeyer R, van Steen ACI, Kempers L, Timmerman AL, Nolte MA, Hombrink P, van Buul JD
Cell reports (2022) 383: 110243. 104 203 WB; tested species: human
Role of SNAREs in the Atopic Dermatitis-related Cytokine Secretion and Skin-Nerve Communication.
Meng J, Wang J, Buddenkotte J, Buhl T, Steinhoff M
The Journal of investigative dermatology (2019) : . 104 203 WB, ICC; KD verified; tested species: human
Adaptor Protein-3-Mediated Trafficking of TLR2 Ligands Controls Specificity of Inflammatory Responses but Not Adaptor Complex Assembly.
Petnicki-Ocwieja T, Kern A, Killpack TL, Bunnell SC, Hu LT
Journal of immunology (Baltimore, Md. : 1950) (2015) 1959: 4331-40. 104 203 WB, ICC
Generation of self-organized autonomic ganglion organoids from fibroblasts.
Liu S, Xiang K, Yuan F, Xiang M
iScience (2023) 263: 106241. 104 203 ICC; tested species: mouse
TLR7 trafficking and signaling in B cells is regulated by the MHCII-associated invariant chain.
Tohme M, Maisonneuve L, Achour K, Dussiot M, Maschalidi S, Manoury B
Journal of cell science (2020) 1335: . 104 203 ICC; tested species: mouse
Role of SNAREs in the Atopic Dermatitis-related Cytokine Secretion and Skin-Nerve Communication.
Meng J, Wang J, Buddenkotte J, Buhl T, Steinhoff M
The Journal of investigative dermatology (2019) : . 104 203 WB, ICC; KD verified; tested species: human
The Kohlschütter-Tönz syndrome associated gene Rogdi encodes a novel presynaptic protein.
Riemann D, Wallrafen R, Dresbach T
Scientific reports (2017) 71: 15791. 104 203 ICC; tested species: rat
Adaptor Protein-3-Mediated Trafficking of TLR2 Ligands Controls Specificity of Inflammatory Responses but Not Adaptor Complex Assembly.
Petnicki-Ocwieja T, Kern A, Killpack TL, Bunnell SC, Hu LT
Journal of immunology (Baltimore, Md. : 1950) (2015) 1959: 4331-40. 104 203 WB, ICC
Botulinum protease-cleaved SNARE fragments induce cytotoxicity in neuroblastoma cells.
Arsenault J, Cuijpers SA, Ferrari E, Niranjan D, Rust A, Leese C, O'Brien JA, Binz T, Davletov B
Journal of neurochemistry (2014) 1295: 781-91. 104 203 ICC; tested species: mouse
Spatial proteomics in neurons at single-protein resolution.
Unterauer EM, Shetab Boushehri S, Jevdokimenko K, Masullo LA, Ganji M, Sograte-Idrissi S, Kowalewski R, Strauss S, Reinhardt SCM, Perovic A, Marr C, et al.
Cell (2024) 1877: 1785-1800.e16. 104 203 DNA-PAINT; tested species: rat
Cat. No.: 104 203
Amount: 50 µg
Price: $455.00
The endothelial diapedesis synapse regulates transcellular migration of human T lymphocytes in a CX3CL1- and SNAP23-dependent manner.
Schoppmeyer R, van Steen ACI, Kempers L, Timmerman AL, Nolte MA, Hombrink P, van Buul JD
Cell reports (2022) 383: 110243. 104 203 WB; tested species: human
Role of SNAREs in the Atopic Dermatitis-related Cytokine Secretion and Skin-Nerve Communication.
Meng J, Wang J, Buddenkotte J, Buhl T, Steinhoff M
The Journal of investigative dermatology (2019) : . 104 203 WB, ICC; KD verified; tested species: human
Adaptor Protein-3-Mediated Trafficking of TLR2 Ligands Controls Specificity of Inflammatory Responses but Not Adaptor Complex Assembly.
Petnicki-Ocwieja T, Kern A, Killpack TL, Bunnell SC, Hu LT
Journal of immunology (Baltimore, Md. : 1950) (2015) 1959: 4331-40. 104 203 WB, ICC
Generation of self-organized autonomic ganglion organoids from fibroblasts.
Liu S, Xiang K, Yuan F, Xiang M
iScience (2023) 263: 106241. 104 203 ICC; tested species: mouse
TLR7 trafficking and signaling in B cells is regulated by the MHCII-associated invariant chain.
Tohme M, Maisonneuve L, Achour K, Dussiot M, Maschalidi S, Manoury B
Journal of cell science (2020) 1335: . 104 203 ICC; tested species: mouse
Role of SNAREs in the Atopic Dermatitis-related Cytokine Secretion and Skin-Nerve Communication.
Meng J, Wang J, Buddenkotte J, Buhl T, Steinhoff M
The Journal of investigative dermatology (2019) : . 104 203 WB, ICC; KD verified; tested species: human
The Kohlschütter-Tönz syndrome associated gene Rogdi encodes a novel presynaptic protein.
Riemann D, Wallrafen R, Dresbach T
Scientific reports (2017) 71: 15791. 104 203 ICC; tested species: rat
Adaptor Protein-3-Mediated Trafficking of TLR2 Ligands Controls Specificity of Inflammatory Responses but Not Adaptor Complex Assembly.
Petnicki-Ocwieja T, Kern A, Killpack TL, Bunnell SC, Hu LT
Journal of immunology (Baltimore, Md. : 1950) (2015) 1959: 4331-40. 104 203 WB, ICC
Botulinum protease-cleaved SNARE fragments induce cytotoxicity in neuroblastoma cells.
Arsenault J, Cuijpers SA, Ferrari E, Niranjan D, Rust A, Leese C, O'Brien JA, Binz T, Davletov B
Journal of neurochemistry (2014) 1295: 781-91. 104 203 ICC; tested species: mouse
Spatial proteomics in neurons at single-protein resolution.
Unterauer EM, Shetab Boushehri S, Jevdokimenko K, Masullo LA, Ganji M, Sograte-Idrissi S, Kowalewski R, Strauss S, Reinhardt SCM, Perovic A, Marr C, et al.
Cell (2024) 1877: 1785-1800.e16. 104 203 DNA-PAINT; tested species: rat
Background

Synaptobrevins/VAMPs represents a family of integral membrane proteins of 11-13 kDa with the N-terminal region exposed to the cytoplasm and a C-terminal transmembrane domain. Two isoforms were identified in the mammalian CNS, synaptobrevin 1 (VAMP 1 or p18-1) and synaptobrevin 2 (VAMP 2 or p18-2) that differ in their distribution within different brain regions.
Synaptobrevin 1 is highly conserved between vertebrates and invertebrates. It is a major constituent of synaptic vesicles and peptidergic secretory granules in all neurons examined so far. In addition, it is present on secretory granules of neuroendocrine cells. Low levels of synaptobrevin 2 are present in many other tissues where the protein resides on specialized microvesicles.
In non-neuronal cells the third isoform, cellubrevin (VAMP 3), is present where it is localized to an endosomal membrane pool.
Synaptobrevin/VAMP is an essential component of the exocytotic fusion machine, related to a larger protein family referred to as v-SNAREs. It is the sole target for tetanus and several of the botulinal neurotoxins which cleave the protein at single sites in the C-terminal portion of the molecule.