Anti-TIA1 antibody [EPR9304] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

Immunocytochemistry/Immunofluorescence analysis of HuT-78 cells labelling TIA1 with purified ab140595 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140595).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling TIA1 with unpurified ab140595 at a dilution of 1/100.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140595).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

This IHC data was generated using the same anti-TIA1 antibody clone, EPR9304, in a different buffer formulation (cat# ab140595).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling TIA1 with purified ab140595 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IP

Lab

Immunoprecipitation - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

ab140595 (purified) at 1/40 immunoprecipitating TIA1 in HuT-78 whole cell lysate.

Lane 1 (input) : HuT-78 whole cell lysate (10µg)

Lane 2 (+) : ab140595 + HuT-78 whole cell lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab140595 in HuT-78 whole cell lysate.

For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab140595).

All lanes:

Immunoprecipitation - Anti-TIA1 antibody [EPR9304] (<a href='/en-us/products/primary-antibodies/tia1-antibody-epr9304-ab140595'>ab140595</a>)

Predicted band size: 43 kDa

Observed band size: 42 kDa,43 kDa

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• IP

Lab

Immunoprecipitation - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

This data was developed using ab140595, the same antibody clone in a different buffer formulation.

Immunoprecipitation of TIA1 in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 1.0 µg of ab263945 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 [EPR9304] (<a href='/en-us/products/primary-antibodies/tia1-antibody-epr9304-ab140595'>ab140595</a>) at 1 µg

All lanes:

HAP1 cells

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• WB

Lab

Western blot - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

This WB data was generated using the same anti-TIA1 antibody clone, EPR9304, in a different buffer formulation (cat# ab140595).

Lane 1 : Wild-type HAP1 cell lysate (40 μg)
Lane 2 : TIA1 knockout HAP1 cell lysate (40 μg)
Lane 3 : Jurkat cell lysate (40 μg)
Lane 4 : K562 cell lysate (40 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab140595 observed at 43 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab140595 was shown to specifically react with TIA1 when TIA1 knockout samples were used. Wild-type and TIA1 knockout samples were subjected to SDS-PAGE. ab140595 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TIA1 antibody [EPR9304] (<a href='/en-us/products/primary-antibodies/tia1-antibody-epr9304-ab140595'>ab140595</a>)

Predicted band size: 43 kDa

false

• WB

Lab

Western blot - Anti-TIA1 antibody [EPR9304] - BSA and Azide free (AB230829)

This data was developed using ab140595, the same antibody clone in a different buffer formulation.

ab140595 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 ab140595 overnight at 4 °C at a 1/5000 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 [EPR9304] (<a href='/en-us/products/primary-antibodies/tia1-antibody-epr9304-ab140595'>ab140595</a>) at 1/5000 dilution

Lane 1:

Wild-type HAP1 lysate at 50 µg

Lane 2:

TIA1 knock-out HAP1 lysate at 50 µg

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