Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

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 - BSA and Azide free
  • Suitable for: Flow Cyt, IP, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free
    See all TIA1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22999-80] to TIA1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IP, WB, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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: WT HAP1 and Jurkat cells. IP: Jurkat and HuT-78 cells.

  • General notes

    ab263953 is the carrier-free version of ab263945. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab263953 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Western blot - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Flow Cytometry - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Flow Cytometry - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Flow Cytometry - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Flow Cytometry - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263945).

  • Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)
    Anti-TIA1 antibody [EPR22999-80] - BSA and Azide free (ab263953)

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