Anti-MTA1 antibody [EPR24507-64] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTA1 antibody [EPR24507-64] - BSA and Azide free (AB288770)

This data was developed using ab288765, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling MTA1 with ab288765 at 1/50 (9.62 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MTA1 antibody [EPR24507-64] - BSA and Azide free (AB288770)

This data was developed using ab288765, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling MTA1 with ab288765 at 1/500 dilution (0.1ug)(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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MTA1 antibody [EPR24507-64] - BSA and Azide free (AB288770)

This data was developed using ab288765, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling MTA1 with ab288765 at 1/50 (9.62 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in 293T cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• WB

Lab

Western blot - Anti-MTA1 antibody [EPR24507-64] - BSA and Azide free (AB288770)

This data was developed using ab288765, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Lanes 1-2 : This blot was developed using a higher sensitivity ECL substrate.

Lysates was made freshly and used in WB immediately.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-MTA1 antibody [EPR24507-64] (<a href='/en-us/products/primary-antibodies/mta1-antibody-epr24507-64-ab288765'>ab288765</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 4:

A549 (Human lung carcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 81 kDa

Observed band size: 78-82 kDa

false

• WB

Lab

Western blot - Anti-MTA1 antibody [EPR24507-64] - BSA and Azide free (AB288770)

This data was developed using the same antibody clone in a different buffer formulation (ab288765).

Western blot : Anti-MTA1 antibody [EPR24507-64] (ab288765) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab288765 was shown to bind specifically to MTA1. A band was observed at 80-85 kDa in wild-type A549 cell lysates with no signal observed at this size in MTA1 knockout cell line. To generate this image, wild-type and MTA1 knockout A549 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.

All lanes:

Western blot - Anti-MTA1 antibody [EPR24507-64] (<a href='/en-us/products/primary-antibodies/mta1-antibody-epr24507-64-ab288765'>ab288765</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lanes 2 and 4:

Empty cell lysate at 20 µg

Lane 3:

MTA1 knockout A549 ab314986 cell lysate at 20 µg

Lane 5:

Wild-type HEK-293 ab259780 cell lysate at 20 µg

Lane 6:

MTA1 knockout HEK-293 <a href='/en-us/products/cell-lines/human-mta1-knockout-hek-293-cell-line-ab277164'>ab277164</a> cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 80 kDa,85 kDa

false