重组Anti-SMCHD1抗体[EPR27952-42] (ab317811)

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

Low expression: MCF7.

The lane 9 of this blot was developed using a high sensitivity ECL substrate.

In Western blot, Anti-Vinculin antibody [EPR8185]  (ab129002) staining at 1/10000 dilution.

Exposure time: Lanes 1-8: 180 seconds; Lane 9: 59 seconds.

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

Low expression: liver.

The lanes 4-5 of this blot was developed using a high sensitivity ECL substrate.

The identity of the lower MW bands in lanes 4-5 are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185]  (ab129002) staining at 1/10000 dilution.

Exposure time: Lanes 1-3: 180 seconds; Lanes 4-5: 103 seconds.

SMCHD1 was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab317811 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317811 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate
Lane 2: ab317811 IP in HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab317811 in HCT 116 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 180 seconds.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ HCT 116 (human colorectal carcinoma epithelial cell) cells and 5 µg of ab317811 [EPR27952-42]. 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 was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ HCT 116 (human colorectal carcinoma epithelial cell) cells and 5 µg of ab317811 [EPR27952-42]. 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 was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ HCT 116 (human colorectal carcinoma epithelial cell) cells and 5 µg of ab317811 [EPR27952-42]. 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.