Anti-Dnmt3b antibody [EPR26716-583]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3b antibody [EPR26716-583] (AB321879)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labelling Dnmt3b with ab321879 at 1/1000 (0.511 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 NCCIT cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : HT-29 (PMID : 22563479).
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

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ChIP-sequencing - Anti-Dnmt3b antibody [EPR26716-583] (AB321879)

Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab321879 [EPR26716-583]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Dnmt3b antibody [EPR26716-583] (AB321879)

Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab321879 [EPR26716-583]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Dnmt3b antibody [EPR26716-583] (AB321879)

Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 μg of ab321879 [EPR26716-583]. 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-Dnmt3b antibody [EPR26716-583] (AB321879)

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

Negative control : HT-29, WI-38 (PMID : 22563479).

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

All lanes:

Western blot - Anti-Dnmt3b antibody [EPR26716-583] (ab321879) at 1/1000 dilution

Lane 1:

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

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

WI-38 (human fetal lung fibroblast) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 95 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Dnmt3b antibody [EPR26716-583] (AB321879)

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

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

The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Dnmt3b antibody [EPR26716-583] (ab321879) at 1/1000 dilution

Lane 1:

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

Lane 2:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 95 kDa,36 kDa

true

Exposure time: 59s