Overview

  • Product name
  • Description
    Rabbit polyclonal to VAMP2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-Fr, ICC/IF, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
    Predicted to work with: Rhesus monkey
  • Immunogen

    Synthetic peptide corresponding to Rat VAMP2 aa 1-18.
    Sequence:

    MSATAATVPPAAPAGEGG


    (Peptide available as ab4958)

Properties

Applications

Our Abpromise guarantee covers the use of ab3347 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Detects a band of approximately 19 kDa (predicted molecular weight: 19 kDa).Can be blocked with VAMP2 peptide (ab4958).
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/1000.
IHC-FoFr Use at an assay dependent concentration. PubMed: 23056393

Target

  • Function
    Involved in the targeting and/or fusion of transport vesicles to their target membrane.
  • Tissue specificity
    Nervous system and skeletal muscle.
  • Sequence similarities
    Belongs to the synaptobrevin family.
    Contains 1 v-SNARE coiled-coil homology domain.
  • Cellular localization
    Cytoplasmic vesicle > secretory vesicle > synaptic vesicle membrane. Cell junction > synapse > synaptosome. Neuronal synaptic vesicles.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ11460 antibody
    • RATVAMPB antibody
    • RATVAMPIR antibody
    • SYB antibody
    • SYB2 antibody
    • Synaptobrevin 2 antibody
    • Synaptobrevin-2 antibody
    • VAMP 2 antibody
    • VAMP-2 antibody
    • Vamp2 antibody
    • VAMP2_HUMAN antibody
    • Vesicle associated membrane protein 2 antibody
    • Vesicle-associated membrane protein 2 (synaptobrevin 2) antibody
    • Vesicle-associated membrane protein 2 antibody
    see all

Images

  • Western blot detection of VAMP2 on rat brain extract using ab3347.

  • VAMP2 antibody (ab3347) used in Immunohistochemistry (rat frozen sections). Sections were fixed with paraformaldehyde. Primary Antibody ab3347 used at 1/1000 incubated for 18 hours at 20°C in PBS-T 0.3%.  Secondary Antibody used anti-IgG Rabbit Conjugation Alexa Fluor® 488 Dilution 1/1000.

    See Abreview

  • Immunohistochemical analysis of rat spinal cord dorsal horn tissue, staining VAMP2 with ab3347 at 1/200 dilution. Fluorescent-conjugated anti-rabbit IgG (1/200) was used as the secondary antibody.

References

This product has been referenced in:
  • Alexander NJ  et al. SNARE Complex-Associated Proteins in the Lateral Amygdala of Macaca mulatta Following Long-Term Ethanol Drinking. Alcohol Clin Exp Res 42:1661-1673 (2018). Read more (PubMed: 29944190) »
  • Kim D  et al. Graphene quantum dots prevent a-synucleinopathy in Parkinson's disease. Nat Nanotechnol N/A:N/A (2018). Read more (PubMed: 29988049) »
See all 27 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Loading amount
2 µg
Gel Running Conditions
Non-reduced Denaturing
Sample
Rat Tissue lysate - whole (hippocampus tissue lysate)
Specification
hippocampus tissue lysate
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Jul 07 2014

Answer

Thank you for your reply and the additional information.

Please let me know your order or PO number. If the antibody was purchased within the last 6 months, our Abpromise guarantee applies and I'd be happy to offer you either a free of charge replacement, a credit or a refund.

Please let me know which option would be best for you.

I look forward to hear back and resolve this case for you.

Read More

Answer

Thank you for contacting us.
I am sorry to hear that the antibody is not working as expected.

I have copied the WB protocol below.
Also,
I have a few more questions in order to understand the issue better:
1) What is the exact problem (e.g. no bands, high background)?
2) What type of samples have you used and how were they prepared?
3) How long did you incubate with the primary antibody?



Western Blot Protocol

Polyacrylamide gel electrophoresis and blottingAdd an appropriate amount of electrophoresis sample buffer (1X = 125mM Tris-HCl pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, and 1% beta-mercaptoethanol) to all samples.
Heat to 95°C for 3-5 minutes.
Load 5-100 ug total protein in a volume that is appropriate for the size of the wells.
Electrophorese proteins for the appropriate time according to the manufacturers instructions.
Transfer proteins from the gel to a suitable membrane (e.g. nitrocellulose, PVDF, etc.) following the manufacturers protocol for transfer.
High molecular weight proteins (>120 kDa) can be wet transferred more efficiently if transfer time is increased and 0.05% SDS is included in the transfer buffer.


BlockingRemove the filter from the transfer apparatus and rinse in PBST/TBST to remove loose acrylamide.
Transferred proteins can be visualized by staining the membrane for 15-30 minutes with Ponceau S.
Remove stain from filter by washing with PBST/TBST.
Place filter into blocking solution.
Block for 30 minutes at 37°C, 1 hour at room temperature, or overnight at 4°C.


Incubation with primary antibodyDecant the blocking buffer and add the antibody, diluted in blocking buffer as suggested in the product description sheet.
Incubate with agitation for 30 minutes at 37°C, 1 hour at room temperature, or overnight at 4°C.


Incubation with secondary antibodyWash for 30 minutes with agitation in wash buffer (TBS or PBS with 0.1% Tween 20), changing the wash buffer every 5 minutes.
Decant the wash solution and add AffiniClear HRP-conjugated secondary antibody, diluted in 5% non-fat dry milk in wash buffer.
Incubate for 30 minutes at 37°C, 1 hour at room temperature, or overnight at 4°C.
Decant the antibody conjugate and wash for 30 minutes with agitation in wash buffer (TBS or PBS with 0.1% Tween 20), changing the wash buffer every 5 minutes.


Substrate incubation (ECL)Decant washing buffer and place the blot in a plastic bag or clean tray containing the development working solution (0.125 ml/cm2) for 1-5 minutes.
Agitate bag or tray to cover the surface of the membrane.
Remove the blot from the bag or tray and place it between two pieces of write-on transparency film.
Smooth over the covered blot to remove air bubbles and excess substrate.
Expose to X-ray film or any sensitive screen. (Check manufacturer's instructions for specific ECL reagents and procedures.)


I look forward to hear back from you and assist you further.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
No

Dr. Karine Thibault

Verified customer

Submitted Apr 14 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (hippocampal neuron)
Specification
hippocampal neuron
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 2%

Dr. Chanchoo Yap

Verified customer

Submitted Nov 27 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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