Anti-KDM6B / JMJD3 antibody [EPR28325-184]

• WB

Supplier Data

Western blot - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (AB324740)

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (ab324740) at 1/1000 dilution

Lane 1:

Untreated SW620 (human colorectal adenocarcinoma epithelial cell) whole cell fresh lysate at 20 µg

Lane 2:

SW620 treated with 100ng/ml IFN gamma for 8 hours whole cell fresh lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 200 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (AB324740)

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

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

All lanes:

Western blot - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (ab324740) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) transfected with scrambled siRNA control whole cell fresh lysate at 20 µg

Lane 2:

K-562 transfected with siRNA specifically targeting KDM6B whole cell fresh lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 200 kDa,124 kDa

true

Exposure time: 92s

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (AB324740)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ SW620 (human colorectal adenocarcinoma epithelial cell) cells treated with TPA(80nM 3h) and 5 g of ab324740 [EPR28325-184]. 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

Supplier Data

ChIC/CUT&RUN sequencing - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (AB324740)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ SW620 (human colorectal adenocarcinoma epithelial cell) cells treated with TPA(80nM 3h) and 5 g of ab324740 [EPR28325-184]. 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

Supplier Data

ChIC/CUT&RUN sequencing - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (AB324740)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ SW620 (human colorectal adenocarcinoma epithelial cell) cells treated with TPA(80nM 3h) and 5 g of ab324740 [EPR28325-184]. 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.