Recombinant Anti-LAT antibody [EPR26091-2] (ab302960)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26091-2] to LAT
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF, IP
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-LAT antibody [EPR26091-2]
See all LAT primary antibodies -
Description
Rabbit monoclonal [EPR26091-2] to LAT -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat
Does not react with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse and rat thymus tissue lysate. EL4 whole cell lysate. IHC-P: Mouse thymus and spleen tissue. Rat spleen tissue. ICC/IF: EL4 cells. Flow Cyt (Intra): EL4 cells. Mouse splenocytes. IP: Mouse thymus tissue lysate.
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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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR26091-2 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab302960 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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WB |
1/1000. Detects a band of approximately 36 kDa.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/50.
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ICC/IF |
1/100.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 36 kDa. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/50. |
ICC/IF
1/100. |
IP
1/30. |
Target
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Function
Required for TCR (T-cell antigen receptor)- and pre-TCR-mediated signaling, both in mature T-cells and during their development. Involved in FCGR3 (low affinity immunoglobulin gamma Fc region receptor III)-mediated signaling in natural killer cells and FCER1 (high affinity immunoglobulin epsilon receptor)-mediated signaling in mast cells. Couples activation of these receptors and their associated kinases with distal intracellular events such as mobilization of intracellular calcium stores, PKC activation, MAPK activation or cytoskeletal reorganization through the recruitment of PLCG1, GRB2, GRAP2, and other signaling molecules. -
Tissue specificity
Expressed in thymus, T-cells, NK cells, mast cells and, at lower levels, in spleen. Present in T-cells but not B-cells (at protein level). -
Post-translational
modificationsPhosphorylated on tyrosines by ZAP-70 upon TCR activation, or by SYK upon other immunoreceptor activation; which leads to the recruitment of multiple signaling molecules. Is one of the most prominently tyrosine-phosphorylated proteins detected following TCR engagement. May be dephosphorylated by PTPRJ.
Palmitoylation of Cys-26 and Cys-29 is required for raft targeting and efficient phosphorylation. -
Cellular localization
Cell membrane. Present in lipid rafts. - Information by UniProt
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Database links
- Entrez Gene: 16797 Mouse
- Entrez Gene: 81511 Rat
- SwissProt: O54957 Mouse
- SwissProt: O70601 Rat
- Unigene: 10280 Mouse
- Unigene: 9773 Rat
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Alternative names
- 36 kDa phospho tyrosine adapter protein antibody
- 36 kDa phospho-tyrosine adapter protein antibody
- 36 kDa phospho-tyrosine adaptor protein antibody
see all
Images
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All lanes : Anti-LAT antibody [EPR26091-2] (ab302960) at 1/1000 dilution
Lane 1 : Mouse thymus tissue lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : EL4 (mouse lymphoma T lymphocyte), whole cell lysate
Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), 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
Observed band size: 36 kDa why is the actual band size different from the predicted?
Exposure time: 59 secondsBlocking / Diluting buffer and concentration: 5% NFDM/TBST
Negative controls: testis (PMID: 9489702), RAW264.7 (PMID: 19948717)
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All lanes : Anti-LAT antibody [EPR26091-2] (ab302960) at 1/1000 dilution
Lane 1 : Rat thymus tissue lysate
Lane 2 : Rat testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 36 kDa why is the actual band size different from the predicted?
Exposure time: 92 secondsBlocking / Diluting buffer and concentration: 5% NFDM/TBST
Negative control: testis(PMID:9489702)
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Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling LAT with ab302960 at 1/100 (5.36 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing membranous and cytoplasmic staining in EL4 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Negative control: RWA 264.7 (PMID: 19948717) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.
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Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling LAT with ab302960 at 1/500 (1.072 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Positive staining on rat spleen. The section was incubated with ab302960 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling LAT with ab302960 at 1/500 (1.072 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Negative control: No staining on mouse testis. The section was incubated with ab302960 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling LAT with ab302960 at 1/500 (1.072 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Positive staining on mouse spleen. The section was incubated with ab302960 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Mouse thymus tissue labeling LAT with ab302960 at 1/500 (1.072 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Positive staining on mouse thymus. The section was incubated with ab302960 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins.
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Flow cytometric (Intracellular) analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / EL4 (mouse lymphoma T lymphocyte, Right) cells labelling LAT with ab302960 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: RAW264.7 (PMID: 19948717).
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Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse splenocytes cells labelling LAT with ab302960 at 1/50 dilution. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647 then fixed with 2% PFA for 10min followed by intracellularly stained with ab172730 (isotype control) or ab302960.
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LAT was immunoprecipitated from 0.35 mg Mouse thymus tissue lysate 10 µg with ab302960 at 1/30 dilution (2µg in 0.35mg lysates). Primary ab302960 used for western blot at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Mouse thymus tissue lysate 10 µg
Lane 2: ab302960 IP in Mouse thymus tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302960 in Mouse thymus tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab302960 has not yet been referenced specifically in any publications.