Recombinant Anti-TIA1 antibody [EPR22999-80] (ab263945)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22999-80] to TIA1
- Suitable for: Flow Cyt (Intra), IP, IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-TIA1 antibody [EPR22999-80]
See all TIA1 primary antibodies -
Description
Rabbit monoclonal [EPR22999-80] to TIA1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, IHC-P, WBmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1, Jurkat, HuT-78, MOLT-4, K562 and Human tonsil lysates. IHC-P: Human spleen, Human classical hodgkin lymphoma and Human gastric cancer tissues. Flow Cyt (intra): WT HAP1 and Jurkat cells. IP: Jurkat and HuT-78 cells.
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22999-80 -
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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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab263945 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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Flow Cyt (Intra) |
1/600.
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IP |
1/30.
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 43 kDa.
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Notes |
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Flow Cyt (Intra)
1/600. |
IP
1/30. |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 43 kDa. |
Target
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Function
Involved in alternative pre-RNA splicing and regulation of mRNA translation by binding to AU-rich elements (AREs) located in mRNA 3' untranslated regions (3' UTRs). Possesses nucleolytic activity against cytotoxic lymphocyte target cells. May be involved in apoptosis. -
Sequence similarities
Contains 3 RRM (RNA recognition motif) domains. -
Cellular localization
Cytoplasmic granule. Nucleus. Accumulates in cytoplasmic stress granules (SG) following cellular damage. - Information by UniProt
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Database links
- Entrez Gene: 7072 Human
- Omim: 603518 Human
- SwissProt: P31483 Human
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Alternative names
- Cytotoxic granule associated RNA binding protein 1 antibody
- Cytotoxic granule associated RNA binding protein antibody
- mTIA-1 antibody
see all
Images
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All lanes : Anti-TIA1 antibody [EPR22999-80] (ab263945) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TIA1 knockout HAP1 whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 4 : HuT-78 (human Sezary syndrome cutaneous T lymphocyte), whole cell lysate
Lane 5 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
Lane 6 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 7 : Human tonsil tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 43 kDa
Observed band size: 42,43 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:30533021).
ab263945 was shown to specifically react with TIA1 in wild-type HAP1 cells as signal was lost in TIA1 knockout cells. Wild-type and TIA1 knockout samples were subjected to SDS-PAGE. ab263945 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
Exposure time: Lanes 1-3: 103 seconds Lane 4: 26 seconds Lanes 5-6: 103 seconds Lane 7: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labeling TIA1 with ab263945 at 1/4000 dilution (0.15ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in tumor-infiltrating T-lymphocytes of human gastric cancer (PMID: 10658910) is observed.
The section was incubated with ab256492 for 15 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized TIA1 knockout HAP1 (TIA1 knockout human chronic myelogenous leukemia near-haploid cell line, (Left) / WT HAP1 (Right) cells labelling TIA1 with ab263945 at 1/600 dilution (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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TIA1 was immunoprecipitated from 0.35 mg HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate with ab263945 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10ug
Lane 2: ab263945 IP in HuT-78 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263945 in HuT-78 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Truncated form (~16kDa) of TIA1 was also immunoprecipitated. (PMID: 25224594).
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TIA1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug with ab263945 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug
Lane 2: ab263945 IP in Jurkat whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263945 in Jurkat whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Truncated form (~16kDa) of TIA1 was also immunoprecipitated. (PMID: 25224594).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labelling TIA1 with ab263945 at 1/600 dilution (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded Human classical hodgkin lymphoma tissue labeling TIA1 with ab263945 at 1/4000 dilution (0.15ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in T-lymphocytes of human classical hodgkin lymphoma (PMID: 19096012) is observed.
The section was incubated with ab256492 for 15 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling TIA1 with ab263945 at 1/4000 dilution (0.15ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in T-lymphocytes of human spleen (PMID: 23510456) is observed.
The section was incubated with ab256492 for 15 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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 (1)
ab263945 has been referenced in 1 publication.
- Chen B et al. Elevated LINC00894 relieves the oncogenic properties of thyroid cancer cell by sponging let-7e-5p to promote TIA-1 expression. Discov Oncol 13:56 (2022). PubMed: 35776220