Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2629Y] to VAMP8/EDB - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free
    See all VAMP8/EDB primary antibodies

  • Description

    Rabbit monoclonal [EP2629Y] to VAMP8/EDB - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Rat
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431, 293T, HEK293, HeLa, RAW264.7, NIH/3T3 and PC-12 cell lysates; Mouse kidney tissue lysate. IHC-P: Human brain and kidney tissue. ICC/IF: PC-12 cells. Flow Cyt: HeLa cells. IP: HEK293 cell lysate.

  • General notes

    ab227984 is the carrier-free version of ab76021 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab227984 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Previously labelled as VAMP8.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Immunocytochemistry/ Immunofluorescence - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

    Immunocytochemistry/Immunofluorescence analysis of PC-12 cells labelling VAMP8/EDB with purified ab76021 at a dilution of 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76021).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling VAMP8/EDB with purified ab76021 at a dilution of 1/250. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76021).

  • Western blot - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Western blot - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    All lanes : Anti-VAMP8/EDB antibody [EP2629Y] (ab76021) at 1/10000 dilution

    Lane 1 : Wild-type A431 cell lysate
    Lane 2 : VAMP8 knockout A431 cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 11 kDa
    Observed band size: 13 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab76021).

    Lanes 1 - 4: Merged signal (red and green). Green - ab76021 observed at 13 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab76021 was shown to react with VAMP8/EDB in wild-type A431 cells in western blot. Loss of signal was observed when VAMP8 knockout sample was used. Wild-type and VAMP8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab76021 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Flow Cytometry - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

    Flow Cytometry analysis of HeLa cells labelling VAMP8/EDB with purified ab76021 at a dilution of 1/150 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76021).

  • Immunoprecipitation - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Immunoprecipitation - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

    ab76021 (purified) at 1/50 immunoprecipitating VAMP8/EDB in HEK293 whole cell lysate.

    Lane 1 (input): HEK293 whole cell lysate (10µg)

    Lane 2 (+): ab76021 + HEK293 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76021 in HEK293 whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76021).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984) Image from Kamoi M et al. PLoS One. 2012;7(9):e43688. doi: 10.1371/journal.pone.0043688. Epub 2012 Sep 4. Fig 4.; doi:10.1371/journal.pone.0043688; September 4 2012 PLoS ONE 7(9): e43688.

    Immunohistochemical analysis of Human lacrimal gland tissue staining VAMP8/EDB with unpurified ab76021.

    Antigen retrieval was performed using antigen retrieval solution in a microwave. Sections were blocked with 10 goat serum for 30 minutes and incubated with primary antibody (1/100) overnight at 4°C. Staining was detected using DAB.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76021).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

    This IHC data was generated using the same anti-VAMP8/EDB antibody clone, EP2629Y, in a different buffer formulation (cat# ab76021).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labelling VAMP8/EDB with unpurified ab76021 at a dilution of 1/100. A HRP/AP polymerized secondary antibody was used.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)
    Anti-VAMP8/EDB antibody [EP2629Y] - BSA and Azide free (ab227984)

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