VAMP2 Recombinant Rabbit Monoclonal Antibody [JM11-00]
Catalog# ET1703-50
VAMP2 Recombinant Rabbit Monoclonal Antibody [JM11-00]
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WB
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IF-Cell
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IHC-P
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FC(Intra)
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IHC-Fr
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
VAMP2 Recombinant Rabbit Monoclonal Antibody [JM11-00]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human VAMP2 aa 9-42 / 116.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IHC-P, FC(Intra), IHC-Fr, IP
Target Molecular Weight
Predicted band size: 13 kDa
Positive Control
Human brain tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate, U-87 MG, Neuro-2a, C6, mouse brain tissue, rat brain tissue.
Conjugation
unconjugated
Clone Number
JM11-00
RRID
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | IHC-Fr | FC(Intra) | IF-Cell | |
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| human |
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| mouse |
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| rat |
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
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1:500
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IHC-P
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1:5,000-1:10,000
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FC(Intra)
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1:1,000
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IHC-Fr
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1:500-1:1,000
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IP
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Use at an assay dependent concentration.
Target
Function
Syntaxins were originally thought to be docking proteins, but have more recently been categorized as anchoring proteins that anchor themselves to the cytoplasmic surfaces of cellular membranes. Syntaxins have been shown to bind to various proteins involved in exocytosis, including VAMPs (vesicle-associated membrane proteins), NSF (N-ethylmaleimide-sensitive factor), SNAP 25 (synaptosomal-associated protein of 25 kDa), SNAPs (soluble NSF attachment proteins) and synaptotagmin. VAMPs, also designated synaptobrevins, including VAMP-1 and VAMP-2, and synaptotagmin, a protein that may function as an inhibitor of exocytosis, are vesicular proteins. SNAPs, including a- and γ-SNAP, are cytoplasmic proteins that bind to a membrane receptor complex composed of VAMP, SNAP 25 and syntaxin. SNAPs mediate the membrane binding of NSF, which is essential for membrane fusion reactions. An additional protein designated synaptophysin may regulate exo-cytosis by competing with SNAP 25 and syntaxins for VAMP binding.
Background References
1. Xu S et al. Autocrine insulin increases plasma membrane KATP channel via PI3K-VAMP2 pathway in MIN6 cells. Biochem Biophys Res Commun 468:752-7 (2015).
2. Kosiorek M et al. Calcineurin/NFAT signaling represses genes Vamp1 and Vamp2 via PMCA-dependent mechanism during dopamine secretion by Pheochromocytoma cells. PLoS One 9:e92176 (2014).
Sequence Similarity
Belongs to the synaptobrevin family.
Tissue Specificity
Nervous system and skeletal muscle.
Post-translational Modification
Phosphorylated by PRKCZ in vitro and this phosphorylation is increased in the presence of WDFY2.; (Microbial infection) Targeted and hydrolyzed by C.botulinum neurotoxin type B (BoNT/B, botB) which hydrolyzes the 76-Gln-|-Phe-77 bond and probably inhibits neurotransmitter release.; (Microbial infection) Targeted and hydrolyzed by C.botulinum neurotoxin type D (BoNT/D, botD) which probably hydrolyzes the 59-Lys-|-Leu-60 bond and inhibits neurotransmitter release. Note that humans are not known to be infected by C.botulinum type D.; (Microbial infection) Targeted and hydrolyzed by C.botulinum neurotoxin type F (BoNT/F, botF) which hydrolyzes the 58-Gln-|-Lys-59 bond and probably inhibits neurotransmitter release.; (Microbial infection) Targeted and hydrolyzed by C.tetani tetanus toxin (tetX) which hydrolyzes the 76-Gln-|-Phe-77 bond and probably inhibits neurotransmitter release.
Subcellular Location
Cell junction, Cell membrane, Cytoplasmic vesicle , Synapse, Synaptosome.
Synonyms
FLJ11460 antibody
RATVAMPB antibody
RATVAMPIR antibody
SYB antibody
SYB2 antibody
Synaptobrevin 2 antibody
Synaptobrevin-2 antibody
VAMP 2 antibody
VAMP-2 antibody
Vamp2 antibody
ExpandImages
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Application: Immunofluorescence (IHC-Fr)
Species: Mouse
Tissue: Retina
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1703-50, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Application: Immunofluorescence (IHC-Fr)
Species: Rat
Tissue: Retina
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1703-50, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Western blot analysis of VAMP2 on different lysates with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/1,000 dilution.
Lane 1: Human brain tissue lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Rat brain tissue lysate
Lysates/proteins at 40 µg/Lane.
Predicted band size: 13 kDa
Observed band size: 18 kDa
Exposure time: 2 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-50) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded mouse retina tissue with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-50) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-50) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-50) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-50) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-50) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Application: Immunofluorescence (IHC-Fr)
Species: Rat
Tissue: Hippocampus
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1703-50, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Immunocytochemistry analysis of U-87 MG cells labeling VAMP2 with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of Neuro-2a cells labeling VAMP2 with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling VAMP2 with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-VAMP2 antibody (ET1703-50) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of U-87 MG cells labeling VAMP2.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-50, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"