Antibodies, Proteins, Kits and Reagents for Life Science

254 630 ₸ Product size

100 µl 254 630 ₸

Order now and get it on Tuesday March 09, 2021

Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP2853Y] to SLP76 (phospho Y145)
Suitable for: ICC/IF, Dot blot, WB, Flow Cyt
Reacts with: Human

Product name

Anti-SLP76 (phospho Y145) antibody [EP2853Y]
See all SLP76 primary antibodies
Description

Rabbit monoclonal [EP2853Y] to SLP76 (phospho Y145)
Host species

Rabbit
Specificity

This antibody detects SLP76 phosphorylated on tyrosine 145.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Jurkat whole cell lysate (ab7899) treated with pervanadate.
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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EP2853Y
Isotype

IgG
Research areas

Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/5000 dilution

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates. treated with pervanadate at 50nM for 5 min.
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates. Treated with pervanadate at 50nM for 5 min. Then the membrane was incubated with phosphatase.

Lysates/proteins at 15 µg per lane.

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

Predicted band size: 60 kDa
Observed band size: 76 kDa
why is the actual band size different from the predicted?

Blocking buffer 5% NFDM/TBST. Diluting buffer 5% NFDM/TBST
Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

ab75829 staining SLP76 in Jurkat by Immunocytochemistry. Tissue was fixed with 4% paraformaldehyde. Samples were incubated with primary antibody (4.5 μg/ml). ab150077 AlexaFluor®488 Goat anti-Rabbit (1/1000) was used as the secondary antibody. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 2.5 μg/ml and DAPI were used as counter stains.

Confocal image showing the expression was increased after treatment with pervanadate (1mM, 30min) and then decreased after treatment with the Lambda Protein Phosphatase 31℃ for 2h
Immunocytochemistry/ Immunofluorescence - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

ab75829 staining SLP76 (phospho Y145) antibody in both treated (50mM of pervanadate for 5 minutes) and untreated Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000.

DAPI was used as a nuclear counterstain.
Western blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/200000 dilution

Lane 1 : Jurkat cell lysates un-treated
Lane 2 : Jurkat cell lysates treated with pervanadate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 60 kDa
Observed band size: 76 kDa why is the actual band size different from the predicted?
Dot Blot - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

Dot blot analysis of SLP76 (phospho Y145) phospho peptide (Lane 1), SLP76 Non-phospho peptide (Lane 2), labelling SLP76 (phospho Y145) with ab75829 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.

Blocking and diluting buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Flow Cytometry - Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

Flow cytometric analysis of permeabilized Jurkat cells, untreated (green) or pervanadate treated (red) using ab75829 at a 1/100 dilution. Also, pervanadate Jurkat cells using the same antibody preincubated with phospho SLP76 peptide (blue) or non phospho SLP76 peptide (orange).
Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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