Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸ 10 µl 130 665 ₸

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Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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
Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-VAMP8/EDB antibody [EP2629Y]
See all VAMP8/EDB primary antibodies
Description

Rabbit monoclonal [EP2629Y] to VAMP8/EDB
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Rat
IHC-P	Human
IP	Human
WB	Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human VAMP8/EDB (N terminal). The exact sequence is proprietary.
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
Previously labelled as VAMP8.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP2629Y
Isotype

IgG
Research areas

Western blot - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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?

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 A431 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021) 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] (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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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.
Immunocytochemistry/ Immunofluorescence - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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).
Flow Cytometry - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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.
Western blot - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

All lanes : Anti-VAMP8/EDB antibody [EP2629Y] (ab76021) at 1/10000 dilution (purified)

Lane 1 : HEK293 whole cell lysate
Lane 2 : HeLa whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 11 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

All lanes : Anti-VAMP8/EDB antibody [EP2629Y] (ab76021) at 1/10000 dilution (purified)

Lane 1 : Rat kidney tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : RAW264.7 whole cell lysate
Lane 4 : NIH/3T3 whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 11 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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.
Western blot - Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

All lanes : Anti-VAMP8/EDB antibody [EP2629Y] (ab76021) at 1/20000 dilution (unpurified)

Lane 1 : 293T cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : PC12 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 11 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Anti-VAMP8/EDB antibody [EP2629Y] (ab76021)

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