Anti-MED12 antibody [BLR084G]

• ICC

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Immunocytochemistry - Anti-MED12 antibody [BLR084G] (AB275962)

HCT-116 cells stained for MED12 using ab275962 at 1/500 dilution in immunocytochemistry.

• Flow Cyt

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Flow Cytometry - Anti-MED12 antibody [BLR084G] (AB275962)

Flow cytometry overlay histogram of 4% formaldehyde fixed Jurkat cells permeabilized with 90% methanol labeling human MED12 with ab275962 at 1 µl per 1 x 10^6 cells (shaded) compared with a Isotype control (unshaded). Secondary antibody details : DyLight® 488-conjugated goat anti-rabbit IgG (ab96883).

• IP

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Immunoprecipitation - Anti-MED12 antibody [BLR084G] (AB275962)

Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer.

ab275962 used for IP at 20 μl/mg lysate. For blotting immunoprecipitated MED12, ab275962 was used at 1/1000 dilution. Chemiluminescence with an exposure time of 10 seconds.

All lanes:

Immunoprecipitation - Anti-MED12 antibody [BLR084G] (ab275962)

Predicted band size: 243 kDa

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• WB

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Western blot - Anti-MED12 antibody [BLR084G] (AB275962)

Chemiluminescence with an exposure time of 75 seconds.

Lower Panel : Rabbit anti-eEF2

All lanes:

Western blot - Anti-MED12 antibody [BLR084G] (ab275962) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

A549 whole cell lysate at 50 µg

Lane 3:

MCF-7 whole cell lysate at 50 µg

Lane 4:

HepG2 whole cell lysate at 50 µg

Lane 5:

K562 whole cell lysate at 50 µg

Lane 6:

RKO whole cell lysate at 50 µg

Lane 7:

TCMK-1 whole cell lysate at 50 µg

Lane 8:

NIH/3T3 whole cell lysate at 50 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG

Predicted band size: 243 kDa

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• WB

Lab

Western blot - Anti-MED12 antibody [BLR084G] (AB275962)

Western blot : Anti-MED12 antibody [BLR084G] (ab275962) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab275962 was shown to bind specifically to MED12. A band was observed at 238-243 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MED12 knockout cell line. To generate this image, wild-type and MED12 knockout HCT 116 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-MED12 antibody [BLR084G] (ab275962) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

MED12 knockout HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human MED12 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-med12-knockout-hct116-cell-line-ab286465'>ab286465</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A549 Membrane cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 238-243 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MED12 antibody [BLR084G] (AB275962)

Paraffin embedded renal carcinoma tissue stained for MED12 using ab275962 at 1/500 dilution in immunohistochemical analysis.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MED12 antibody [BLR084G] (AB275962)

Paraffin embedded breast carcinoma tissue stained for MED12 using ab275962 at 1/500 dilution in immunohistochemical analysis.