Anti-SMARCA2 / BRM antibody [EPR28611-92]

• IP

Supplier Data

Immunoprecipitation - Anti-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

SMARCA2 / BRM was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab323179 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323179 at 1000 dilution.

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied for Immunoprecipitation as soon as possible.

All lanes:

Immunoprecipitation - Anti-SMARCA2 / BRM antibody [EPR28611-92] (ab323179) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab323179 at 1/30 IP in HeLa (human cervical adenocarcinoma epithelial 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 ab323179 in HeLa 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: 200 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

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 (200 kDa) are likely to be degradation products. In lane 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-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SMARCA2 / BRM antibody [EPR28611-92] (ab323179) at 1/1000 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) 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: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

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

Low expression : SW-13, NCCIT (PMID : 16007216).

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-SMARCA2 / BRM antibody [EPR28611-92] (ab323179) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 50 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

SW-13 (human adrenal gland cortex epithelial cell) whole cell lysate at 50 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

false

Exposure time: 81s

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ HeLa (human cervix adenocarcinoma epithelial cell) cells and 5 µg of ab323179[EPR28611-92]. 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-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ HeLa (human cervix adenocarcinoma epithelial cell) cells and 5 µg of ab323179[EPR28611-92]. 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-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ HeLa (human cervix adenocarcinoma epithelial cell) cells and 5 µg of ab323179[EPR28611-92]. 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.

• WB

Supplier Data

Western blot - Anti-SMARCA2 / BRM antibody [EPR28611-92] (AB323179)

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

This antibody does not cross-react with human BRG1.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-SMARCA2 / BRM antibody [EPR28611-92] (ab323179) at 1/1000 dilution

Lane 1:

His-tagged human SMARCA2/BRM fragment at 10 ng

Lane 2:

His-tagged human BRG1 fragment at 10 ng

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: 28 kDa,36 kDa,20-36 kDa

false

Exposure time: 8s