Antibodies, Proteins, Kits and Reagents for Life Science

241 228 ₸ Product size

100 µl 241 228 ₸ 10 µl 97 161 ₸

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Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83)
Suitable for: Flow Cyt, ICC/IF, Dot blot, WB, IP
Reacts with: Human

Product name

Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y]
See all CD3 zeta primary antibodies
Description

Rabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83)
Host species

Rabbit
Specificity

This antibody detects CD3 zeta phosphorylated on tyrosine 83.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human CD3 zeta aa 50-150 (phospho Y83). The exact sequence is proprietary.
Database link: P20963
Positive control
- WB: Jurkat whole cell lysate (ab7899).
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.

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: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP776(2)Y
Isotype

IgG
Research areas

Flow Cytometry - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

Flow Cytometry analysis of Jurkat (human acute T cell leukemia) treated (Red)/untreated (Green) with 1mM pervanadate for 4 hours with purified ab68236 at 1/250 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

All lanes : Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236) at 1/2000 dilution

Lane 1 : Untreated Jurkat cells whole cell lysates
Lane 2 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates
Lane 3 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 18 kDa
Observed band size: 18 kDa

Exposure time: 3 minutes

Blocking buffer 5% NFDM/TBST

Diluting buffer 5% NFDM/TBST
Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

All lanes : Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236) at 1/10000 dilution

Lane 1 : Jurkat cell lysate, untreated.
Lane 2 : Jurkat cell lysate, treated with pervanadate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 18 kDa
Observed band size: 18-22 kDa
why is the actual band size different from the predicted?
Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

Immunocytochemistry/Immunofluorescence analysis of Jurkat cells (untreated, Per treated and Per+LP treated) labelling CD3 zeta (phospho Y83) with ab68236 (left) and CD3 zeta with ab40804 (right) both at a dilution of 1/200. Cells were fixed with 4% paraformaldehyde 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.

The image shows increased cytoplamic staining after Pervanadate (1 mM, 30 min) treatment on Jurkat cells. The LP treatment decreased the cytoplasmic staining caused by Pervanadate.

ab40804 was used as a Pan control for ab68236. The results showed cytoplamic staining on untreated, pervanadate (1 mM, 30 min) treated and Per+LP treated Jurkat cells.
Dot Blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

Dot blot analysis of CD3 zeta (pY83) phospho peptide (lane 1) and CD3 zeta non-phospho peptide (lane 2) labelling CD3 zeta (phospho Y83) with ab68236 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

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