Anti-TIA1 antibody [EPR22999-80]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TIA1 antibody [EPR22999-80] (AB263945)

ab263945 was shown to react with TIA1 in wild-type HAP1 cells in immunocytochemistry with loss of signal observed in a TIA1 knockout cell line. ab263945 at 1/600 dilution.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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.

• IP

Unknown

Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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).

All lanes:

Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] (ab263945)

Predicted band size: 43 kDa

Observed band size: 16 kDa,42 kDa,43 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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).

All lanes:

Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] (ab263945)

Predicted band size: 43 kDa

Observed band size: 16 kDa,42 kDa,43 kDa

false

• IP

Lab

Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] (AB263945)

Immunoprecipitation of TIA1 in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 1.0 µg of 86050 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Immunoprecipitation - Anti-TIA1 antibody [EPR22999-80] (ab263945) at 1 µg

All lanes:

HAP1 cells

false

• WB

Lab

Western blot - Anti-TIA1 antibody [EPR22999-80] (AB263945)

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.

All lanes:

Western blot - Anti-TIA1 antibody [EPR22999-80] (ab263945) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

TIA1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 4:

HuT-78 (human Sezary syndrome cutaneous T lymphocyte), whole cell lysate at 20 µg

Lane 5:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 20 µg

Lane 6:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 7:

Human tonsil tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 43 kDa

Observed band size: 42 kDa,43 kDa

false

• WB

Lab

Western blot - Anti-TIA1 antibody [EPR22999-80] (AB263945)

ab263945 was shown to react with TIA1 in wild-type HAP1 cells in Western blot with loss of signal observed in a TIA1 knockout cell line. Wild-type HAP1 and TIA1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab263945 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-TIA1 antibody [EPR22999-80] (ab263945) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 50 µg

Lane 2:

TIA1 knock-out HAP1 lysate at 50 µg

false