Anti-Chd1 antibody [EPR28727-78]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Chd1 antibody [EPR28727-78] (AB318138)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized CHD1 konckout HEK293T (Right) / Parental HEK293T(EDWT04) (Left) cells labelling Chd1 with ab318138 at 1/500 dilution (0.1ug) /Magenta (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Chd1 antibody [EPR28727-78] (AB318138)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CHD1 KO HEK293T (CHD1 knockout human embryonic kidney epithelial cell) ab266512 cells labelling Chd1 with ab318138 at 1/50 (10.58 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing nuclear staining in wildtype HEK-293T cells (shown in green), and negative staining in CHD1 knockout HEK-293T cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
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 2.5ug/ml dilution (Magenta). 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 2 ug/ml dilution.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Chd1 antibody [EPR28727-78] (AB318138)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab318138 [EPR28727-78]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Chd1 antibody [EPR28727-78] (AB318138)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab318138 [EPR28727-78]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Chd1 antibody [EPR28727-78] (AB318138)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab318138 [EPR28727-78]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• WB

Supplier Data

Western blot - Anti-Chd1 antibody [EPR28727-78] (AB318138)

In Western blot, ab318138 was shown to bind specifically to Chd1. Target of interest was observed at 250 kDa in wild-type HEK-293T cell lysates (lane 1) with no signal observed at this size in CHD1 knockout cell line ab266512 (knockout cell lysate 263151) (lane 2).

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

All lanes:

Western blot - Anti-Chd1 antibody [EPR28727-78] (ab318138) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

CHD1 knockout HEK-293T whole cell lysate at 20 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast) 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: 250 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Chd1 antibody [EPR28727-78] (AB318138)

This antibody does not cross-react with human Chd2.

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

All lanes:

Western blot - Anti-Chd1 antibody [EPR28727-78] (ab318138) at 1/1000 dilution

Lane 1:

His-tagged human Chd1 protein at 10 ng

Lane 2:

His-tagged human Chd2 protein 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: 25 kDa

false

Exposure time: 180s