Anti-KDM6A / UTX antibody [EPR26387-105]

• IP

Supplier Data

Immunoprecipitation - Anti-KDM6A / UTX antibody [EPR26387-105] (AB323566)

KDM6A / UTX was immunoprecipitated from 0.35 mg parental HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate with ab323566 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323566 at 1/1000 dilution.

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

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

All lanes:

Immunoprecipitation - Anti-KDM6A / UTX antibody [EPR26387-105] (ab323566) at 1/1000 dilution

Lane 1:

Parental HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 10 µg

Lane 2:

ab323566 at 1/30 IP in Parental HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323566 in parental HAP1(HOWT01) whole cell lysate at 10 µg

Lane 4:

KDM6A Knockout HAP1 whole cell lysate at 10 µg

Lane 5:

ab323566 at 1/30 IP in KDM6A Knockout HAP1 whole cell lysate at 10 µg

Lane 6:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323566 in KDM6A Knockout HAP1(HO010404) whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 154 kDa

false

Exposure time: 76s

• IP

Supplier Data

Immunoprecipitation - Anti-KDM6A / UTX antibody [EPR26387-105] (AB323566)

KDM6A / UTX was immunoprecipitated from 0.35 mg C2C12 (mouse myoblast) whole cell lysate with ab323566 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323566 at 1/1000 dilution.

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

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

All lanes:

Immunoprecipitation - Anti-KDM6A / UTX antibody [EPR26387-105] (ab323566) at 1/1000 dilution

Lane 1:

C2C12 (mouse myoblast) whole cell lysate at 10 µg

Lane 2:

ab323566 at 1/30 IP in C2C12 (mouse myoblast) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323566 in C2C12 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 154 kDa

false

Exposure time: 50s

• WB

Lab

Western blot - Anti-KDM6A / UTX antibody [EPR26387-105] (AB323566)

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

In lane 1, the lysate was stored at -80℃ prior to Western Blotting. The bands beneath the target band (154 kDa) are likey to be degradation products. In lanes 2, 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-KDM6A / UTX antibody [EPR26387-105] (ab323566) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell frozen lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) 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: 154 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-KDM6A / UTX antibody [EPR26387-105] (AB323566)

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

In Western blot, ab323566 was shown to bind specifically to KDM6A. Target of interest was observed at 154 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in KDM6A knockout cell line (lane 2).

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

All lanes:

Western blot - Anti-KDM6A / UTX antibody [EPR26387-105] (ab323566) at 1/1000 dilution

Lane 1:

Parental HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

KDM6A knockout HAP1 whole cell lysate at 20 µg

Lane 3:

C2C12 (mouse myoblast) whole cell 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: 154 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-KDM6A / UTX antibody [EPR26387-105] (AB323566)

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-KDM6A / UTX antibody [EPR26387-105] (ab323566) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell 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: 154 kDa,36 kDa

false

Exposure time: 180s

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-KDM6A / UTX antibody [EPR26387-105] (AB323566)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ MCF7 (human breast adenocarcinoma epithelial cell) cells and 5 ug of ab323566 [EPR26387-105]. 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-KDM6A / UTX antibody [EPR26387-105] (AB323566)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ MCF7 (human breast adenocarcinoma epithelial cell) cells and 5 ug of ab323566 [EPR26387-105]. 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-KDM6A / UTX antibody [EPR26387-105] (AB323566)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ MCF7 (human breast adenocarcinoma epithelial cell) cells and 5 ug of ab323566 [EPR26387-105]. 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-KDM6A / UTX antibody [EPR26387-105] (AB323566)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ C2C12 (mouse myoblast) cells and 5 ug of ab323566 [EPR26387-105]. 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-KDM6A / UTX antibody [EPR26387-105] (AB323566)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ C2C12 (mouse myoblast) cells and 5 ug of ab323566 [EPR26387-105]. 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-KDM6A / UTX antibody [EPR26387-105] (AB323566)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ C2C12 (mouse myoblast) cells and 5 ug of ab323566 [EPR26387-105]. 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.