Anti-TRIM56 antibody [EPR10583] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (AB249161)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer tissue sections labeling TRIM56 with purified ab154862 at 1/1000 dilution (0.691 μg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154862).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (AB249161)

Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154862).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (AB249161)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154862).

• WB

Lab

Western blot - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (AB249161)

This data was developed using ab154862, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267062 (knockout cell lysate ab258249) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TRIM56 antibody [EPR10583] (<a href='/en-us/products/primary-antibodies/trim56-antibody-epr10583-ab154862'>ab154862</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TRIM56 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-trim56-knockout-a549-cell-line-ab267062'>ab267062</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 81 kDa

Observed band size: 88 kDa

false

• WB

Supplier Data

Western blot - Anti-TRIM56 antibody [EPR10583] - BSA and Azide free (AB249161)

This data was developed using ab154862, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.

ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267063 (knockout cell lysate ab258250) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TRIM56 antibody [EPR10583] (<a href='/en-us/products/primary-antibodies/trim56-antibody-epr10583-ab154862'>ab154862</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TRIM56 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-trim56-knockout-a549-cell-line-ab267063'>ab267063</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 81 kDa

Observed band size: 88 kDa

false