Anti-BAF57/SMARCE1 antibody [EPR8848] - BSA and Azide free

• ChIP

Unknown

ChIP - Anti-BAF57/SMARCE1 antibody [EPR8848] - BSA and Azide free (AB248408)

Chromatin was prepared from HeLa cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab131328 (red), and 20 μl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131328).

• WB

Lab

Western blot - Anti-BAF57/SMARCE1 antibody [EPR8848] - BSA and Azide free (AB248408)

This data was developed using the same antibody clone in a different buffer formulation (ab131328).

Lanes 1-3 : Merged signal (red and green). Green - ab131328 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa.

ab131328 Anti-BAF57/SMARCE1 antibody [EPR8848] - ChIP Grade was shown to specifically react with BAF57/SMARCE1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265780 (knockout cell lysate ab258200) was used. Wild-type and BAF57/SMARCE1 knockout samples were subjected to SDS-PAGE. ab131328 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-BAF57/SMARCE1 antibody [EPR8848] (<a href='/en-us/products/primary-antibodies/baf57-smarce1-antibody-epr8848-ab131328'>ab131328</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SMARCE1 knockout HeLa cell lysate at 20 µg

Lane 3:

MCF7 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: 47 kDa

Observed band size: 51 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-BAF57/SMARCE1 antibody [EPR8848] - BSA and Azide free (AB248408)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (,ab131328).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 MCF7 (human breast adenocarcinoma epithelial cell) cells and 5 µg of ab131328 [EPR8848]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-BAF57/SMARCE1 antibody [EPR8848] - BSA and Azide free (AB248408)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (,ab131328).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 MCF7 (human breast adenocarcinoma epithelial cell) cells and 5 µg of ab131328 [EPR8848]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-BAF57/SMARCE1 antibody [EPR8848] - BSA and Azide free (AB248408)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (,ab131328).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 MCF7 (human breast adenocarcinoma epithelial cell) cells and 5 µg of ab131328 [EPR8848]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.