Anti-CHD6 抗体 [EPR26714-1] (ab302956)

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

The bands beneath the target band (315 kDa) are likely to be degraded target fragments.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

The bands beneath the target band (315 kDa) are likely to be degraded target fragments.

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling CHD6 with ab302956 at 1/50 (10.04 µg/ml) dilution followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Nuclear staining on human cerebrum. The section was incubated with ab302956 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CHD6 with ab302956 at 1/50 (10.04 µg/ml) dilution followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Nuclear staining on human breast (PMID: 22569290). The section was incubated with ab302956 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling CHD6 with ab302956 at 1/50 (10.04 µg/ml) dilution followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Nuclear staining on human breast cancer (PMID: 22569290). The section was incubated with ab302956 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval solution 2) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling CHD6 with ab302956 at 1/50 (10.04 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2µg/ml) dilution (Green). Confocal image showing nuclear staining in HeLa cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2µg/ml) dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling CHD6 with ab302956 at 1/50 dilution (1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

CHD6 was immunoprecipitated from 0.35 mg Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug with ab302956 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302956 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2: ab302956 IP in Hela whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302956 in Hela whole cell lysate

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

Exposure time: 180 seconds

Hela whole cell lysate incubating with ab302956 was used to pulldown CHD6. For WB, detection primary antibody is ab259858.

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