Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free)

• IP

Collaborator

Immunoprecipitation - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (AB302563)

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

Immunoprecipitation of TICAM1 in U-87 MG cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab302562 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was 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-TRIF (TICAM1) antibody [EPR24995-152] (<a href='/en-us/products/primary-antibodies/trif-ticam1-antibody-epr24995-152-ab302562'>ab302562</a>) at 2 µg

false

• IP

Supplier Data

Immunoprecipitation - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (AB302563)

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

TRIF (TICAM1) was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma b lymphocyte) whole cell lysate with ab302562 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302562 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-TRIF (TICAM1) antibody [EPR24995-152] (<a href='/en-us/products/primary-antibodies/trif-ticam1-antibody-epr24995-152-ab302562'>ab302562</a>) at 1/30 dilution

Lane 1:

Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate

Lane 2:

<a href='/en-us/products/primary-antibodies/trif-ticam1-antibody-epr24995-152-ab302562'>ab302562</a> IP in Raji whole cell lysate

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (AB302563)

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

Diluting and blocking buffer and concentration : 5% NFDM/TBST.

Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

The identity of the lower MW band at approximately 18 kDa is unknown.

All lanes:

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (<a href='/en-us/products/primary-antibodies/trif-ticam1-antibody-epr24995-152-ab302562'>ab302562</a>) at 1/1000 dilution

Lane 1:

Ramos (human Burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell 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 1/20000 dilution

Predicted band size: 76 kDa

Observed band size: 98 kDa

false

Exposure time: 3min

• WB

Collaborator

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (AB302563)

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

ab302562 was shown to react with TICAM1 in wild-type U-87 MG cells in Western blot with loss of signal observed in TICAM1 knockout cell line ab305279. Wild-type U-87 MG and TICAM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab302562 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was 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 at 1/1000 dilution

Lane 1:

Wild-type U-87 MG lysate at 30 µg

Lane 2:

Western blot - Human TICAM1 knockout U-87MG cell line (<a href='/en-us/products/cell-lines/human-ticam1-knockout-u-87mg-cell-line-ab305279'>ab305279</a>) at 30 µg

Lane 2:

TICAM1 knock-out U-87 MG lysate at 30 µg

false

• WB

Lab

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (AB302563)

Western blot : Anti-TICAM1 antibody [EPR24995-152] (ab302562) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab302562 was shown to bind specifically to TICAM1. A band was observed at 85 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in TICAM1 knockout cell line. To generate this image, wild-type and TICAM1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. This data was developed using ab302562, the same antibody clone in a different buffer formulation.

Lanes 1 - 4:

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (<a href='/en-us/products/primary-antibodies/trif-ticam1-antibody-epr24995-152-ab302562'>ab302562</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (ab302563)

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

TICAM1 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HL-60 cell lysate at 20 µg

Observed band size: 85 kDa

false

• WB

Supplier Data

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (BSA and Azide free) (AB302563)

This data was developed using ab302562, the same antibody clone in a different buffer formulation.

Diluting and blocking buffer and concentration : 5% NFDM/TBST.

Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

Negative control : HL-60.

All lanes:

Western blot - Anti-TRIF (TICAM1) antibody [EPR24995-152] (<a href='/en-us/products/primary-antibodies/trif-ticam1-antibody-epr24995-152-ab302562'>ab302562</a>) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 2:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell 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: 76 kDa

Observed band size: 98 kDa

false

Exposure time: 59s