Anti-Dnmt1 antibody [60B1220.1]

• WB

Supplier Data

Western blot - Anti-Dnmt1 antibody [60B1220.1] (AB13537)

All lanes:

Western blot - Anti-Dnmt1 antibody [60B1220.1] (ab13537) at 1/1000 dilution

All lanes:

H1299 (Human non-small cell lung carcinoma cell line) lysate

Predicted band size: 183 kDa

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• Flow Cyt

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Flow Cytometry - Anti-Dnmt1 antibody [60B1220.1] (AB13537)

Overlay histogram showing HeLa cells stained with ab13537 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13537, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [60B1220.1] (AB13537)

Immunohistochemistry analysis of formalin-fixed human colon carcinoma tissue labelling Dnmt1 with ab13537 at 1/10000 dilution (12 hours at 4°C), followed by biotin Goat Anti-Mouse antibody at 1/2000 for 1 hour at RT. Counterstain : Hematoxylin (purple/blue) nuclear stain at 200μl for 2 mins at RT.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt1 antibody [60B1220.1] (AB13537)

Immunohistochemistry analysis of mouse medullar kidney tissue labelling Dnmt1 with ab13537 at 1/10000 dilution, followed by biotin Goat Anti-Mouse antibody (brown). Counterstain : Hematoxylin (purple/blue) nuclear stain.

• WB

AbReview6936****

Western blot - Anti-Dnmt1 antibody [60B1220.1] (AB13537)

All lanes:

Western blot - Anti-Dnmt1 antibody [60B1220.1] (ab13537) at 1 µg/mL

Lane 1:

HCT116 whole cell lysate

Lane 2:

HCT116 DNMT1 KO whole cell lysate

Lane 3:

HCT116 DNMT3b KO whole cell lysate

Secondary

All lanes:

HRP conjugated Sheep anti-mouse IgG

Predicted band size: 183 kDa

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Exposure time: 1min

This image is courtesy of an anonymous Abreview

• WB

CiteAb

Western blot - Anti-Dnmt1 antibody [60B1220.1] (AB13537)

Western Blotting using Anti-Dnmt1 antibody [60B1220.1], ab13537. Publication image from Park, Y. J. et al., 2015, Nat Commun, 26139044. Legend direct from paper.

DNMT1 regulates the DNA methylation of the adiponectin promoter R2.(a) Dnmt1 mRNA levels in adipocytes from NCD- (n=4) or HFD-fed (n=3) mice and in adipocytes from WT (n=4) or db/db mice (n=4). (b) Correlation between body mass index and DNMT1 mRNA levels in human adipocytes. mRNA levels were measured by qPCR. r2 and P values are indicated on the graph. (c–e) DNMT1 was suppressed by small interfering RNA in 3T3-L1 cells (n=3). (c) Dnmt1 and adiponectin mRNA levels. mRNA levels were measured by qPCR. (d) Adiponectin protein levels were determined by western blot analysis. (e) Bisulfite sequencing data at the R2. (f–h) DNMT1 overexpression in differentiated 3T3-L1 cells (n=3). (f,g) Dnmt1 and adiponectin mRNA and protein levels were measured by qPCR and western blot analysis. (h) Degree of R2 DNA methylation was examined by bisulfite sequencing. (i) AluI restriction sites in R2. Red and grey arrows indicate the AluI restriction sites and CpG locations in the R2, respectively. Double headed arrow points to PCR amplified region. (j) Restriction enzyme accessibility assay in adipocytes from HFD-fed (n=5) or NCD-fed (n=5) mice. EcoRI and BamHI, which do not digest R2, were used as negative controls. Results are expressed as the mean±s.e.m. Similar results were obtained at least more than three independent experiments. *P<0.05; **P<0.01; ***P<0.001 in a two-tailed Student's t-test. siNC; negative control siRNA. See Supplementary Fig. 11 for full-length images of blots.

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