Recombinant 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 (Intra)
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-SLP76 (phospho Y145) antibody [EP2853Y]
See all SLP76 primary antibodies -
Description
Rabbit monoclonal [EP2853Y] to SLP76 (phospho Y145) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Dot blot, WB, Flow Cyt (Intra)more details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Jurkat whole cell lysate (ab7899) treated with pervanadate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2853Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75829 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 |
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ICC/IF |
1/500.
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Dot blot |
1/1000.
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 60 kDa.
|
Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
1/500. |
Dot blot
1/1000. |
WB
1/1000 - 1/10000. Predicted molecular weight: 60 kDa. |
Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Involved in T-cell antigen receptor mediated signaling. -
Tissue specificity
Highly expressed in spleen, thymus and peripheral blood leukocytes. Highly expressed also in T-cell and monocytic cell lines, expressed at lower level in B-cell lines. Not detected in fibroblast or neuroblastoma cell lines. -
Sequence similarities
Contains 1 SAM (sterile alpha motif) domain.
Contains 1 SH2 domain. -
Domain
The SH2 domain mediates interaction with SHB. -
Post-translational
modificationsPhosphorylated after T-cell receptor activation by ZAP-70. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 3937 Human
- Omim: 601603 Human
- SwissProt: Q13094 Human
- Unigene: 304475 Human
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Alternative names
- 76 kDa tyrosine phosphoprotein antibody
- CG8697 antibody
- LCP 2 antibody
see all
Images
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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
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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
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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.
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All lanes : Anti-SLP76 (phospho Y145) antibody [EP2853Y] (ab75829) at 1/200000 dilution
Lane 1 : Jurkat cell lysate un-treated
Lane 2 : Jurkat cell lysate 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 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.
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Intracellular 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).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab75829 has been referenced in 7 publications.
- Rossatti P et al. Rapid increase in transferrin receptor recycling promotes adhesion during T cell activation. BMC Biol 20:189 (2022). PubMed: 36002835
- Zeng L et al. PLCγ1 promotes phase separation of T cell signaling components. J Cell Biol 220:N/A (2021). PubMed: 33929486
- Zhou J et al. Identification and Characterization of the Amphioxus Lck and Its Associated Tyrosine Phosphorylation-Dependent Inhibitory LRR Receptor. Front Immunol 12:656366 (2021). PubMed: 34149695
- Dong R et al. Rewired signaling network in T cells expressing the chimeric antigen receptor (CAR). EMBO J 39:e104730 (2020). PubMed: 32643825
- Wang QL et al. T Cell Receptor (TCR)-Induced PLC-?1 Sumoylation via PIASxß and PIAS3 SUMO E3 Ligases Regulates the Microcluster Assembly and Physiological Function of PLC-?1. Front Immunol 10:314 (2019). PubMed: 30873169
- Yang CY et al. Dual-specificity phosphatase 14 (DUSP14/MKP6) negatively regulates TCR signaling by inhibiting TAB1 activation. J Immunol 192:1547-57 (2014). PubMed: 24403530
- Gill T & Levine AD Mitochondria-derived hydrogen peroxide selectively enhances T cell receptor-initiated signal transduction. J Biol Chem 288:26246-55 (2013). PubMed: 23880762