Anti-MeCP2 antibody - ChIP Grade

• ChIP

Supplier Data

ChIP - Anti-MeCP2 antibody - ChIP Grade (AB195393)

ChIP assays were performed using human osteosarcoma (U2OS) cells, ab195393 and optimized PCR primer sets. Sheared chromatin from 1x10⁶ cells and 5 μg of antibody were used per ChIP experiment. IgG (1 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the ZMYND10 gene (used as a positive control) and CDC6 gene (used as a negative control). Image shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

• WB

Lab

Western blot - Anti-MeCP2 antibody - ChIP Grade (AB195393)

Western blot : Rabbit Polyclonal to MeCP2 ab195393 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 70 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in MECP2 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MeCP2 antibody - ChIP Grade (ab195393) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Western blot - Human MECP2 knockout U-87 MG cell line (ab306725) at 20 µg

Lane 3:

Wild-type HAP1 at 20 µg

Lane 4:

MECP2 knockout HAP1 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 70 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-MeCP2 antibody - ChIP Grade (AB195393)

TBS-Tween containing 5% skimmed milk

All lanes:

Western blot - Anti-MeCP2 antibody - ChIP Grade (ab195393) at 1/1000 dilution

All lanes:

nuclear extract from HeLa cells at 40 µg

Predicted band size: 52 kDa

false

• WB

Lab

Western blot - Anti-MeCP2 antibody - ChIP Grade (AB195393)

Western blot : Rabbit Polyclonal to MeCP2 ab195393 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 52 kDa in Wild-type HCT 116 ab288559 cell lysates with no signal observed at this size in MECP2 knockout HCT 116 ab289193 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MeCP2 antibody - ChIP Grade (ab195393) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 ab288559 at 20 µg

Lane 2:

Western blot - Human MECP2 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-mecp2-knockout-hct116-cell-line-ab289193'>ab289193</a>) at 20 µg

Lane 3:

Wild-type HAP1 at 20 µg

Lane 4:

MECP2 knockout HAP1 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 52 kDa

false