- PA5-30581 - Invitrogen Antibodies DNMT1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of DNMT1 (red) in human melanocytes. Cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.2% Triton X-100 in PBS for 10 minutes at room temperature, and blocked with 5% normal goat serum for 1 hour at room temperature. Cells were probed with a DNMT1 polyclonal antibody (Product # PA5-30581) at a dilution of 1:500 for 1 hour at room temperature, washed with PBS, and incubated with an anti-rabbit Alexa-Fluor 555-conjugated secondary antibody at a dilution of 1:500 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a fluorescent microscope at 20X magnification. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of DNMT1 (red) in human melanocytes. Cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.2% Triton X-100 in PBS for 10 minutes at room temperature, and blocked with 5% normal goat serum for 1 hour at room temperature. Cells were probed with a DNMT1 polyclonal antibody (Product # PA5-30581) at a dilution of 1:500 for 1 hour at room temperature, washed with PBS, and incubated with an anti-rabbit Alexa-Fluor 555-conjugated secondary antibody at a dilution of 1:500 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a fluorescent microscope at 20X magnification. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of DNMT1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: DNMT1 Polyclonal Antibody (Product # PA5-30581) diluted at 1:2000. Red: phalloidin, a cytoskeleton marker.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of DNMT1 was performed using 70% confluent log phase K-562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with DNMT1 Polyclonal Antibody (Product # PA5-30581) at 1:500 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant nuclear localization. Panel e represents merged image of K-562 cells treated with 5 µM Aza deoxycytidine for 3 days (medium and drug were replaced every 24 hours). The signal is significantly lower in panel e due to down-regulation of DNMT1 expression upon Aza deoxycytidine treatment. Panel f represents untreated control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of DNMT1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: DNMT1 Polyclonal Antibody (Product # PA5-30581) diluted at 1:2000. Red: phalloidin, a cytoskeleton marker.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of DNMT1 was performed using 70% confluent log phase K-562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with DNMT1 Polyclonal Antibody (Product # PA5-30581) at 1:500 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominant nuclear localization. Panel e represents merged image of K-562 cells treated with 5 µM Aza deoxycytidine for 3 days (medium and drug were replaced every 24 hours). The signal is significantly lower in panel e due to down-regulation of DNMT1 expression upon Aza deoxycytidine treatment. Panel f represents untreated control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of DNMT1 (red) in human melanocytes. Cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.2% Triton X-100 in PBS for 10 minutes at room temperature, and blocked with 5% normal goat serum for 1 hour at room temperature. Cells were probed with a DNMT1 polyclonal antibody (Product # PA5-30581) at a dilution of 1:500 for 1 hour at room temperature, washed with PBS, and incubated with an anti-rabbit Alexa-Fluor 555-conjugated secondary antibody at a dilution of 1:500 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a fluorescent microscope at 20X magnification. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: DNMT1 Polyclonal Antibody immunoprecipitates DNMT1 protein in IP experiments. IP samples: HeLa nuclear extract. A. Control with 4 µg of preimmune Rabbit IgG. B. Immunoprecipitation of DNMT1 protein by 4 µg DNMT1 Polyclonal Antibody (Product # PA5-30581). 5 % SDS-PAGE. The immunoprecipitated DNMT1 protein was detected by DNMT1 Polyclonal Antibody (Product # PA5-30581) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: DNMT1 Polyclonal Antibody detects DNMT1 protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse testis. DNMT1 stained by DNMT1 Polyclonal Antibody (Product # PA5-30581) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: DNMT1 Polyclonal Antibody detects DNMT1 protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded human tonsil. DNMT1 stained by DNMT1 Polyclonal Antibody (Product # PA5-30581) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of DNMT1 was performed in paraffin-embedded human endometrial cancer tissue using DNMT1 Polyclonal Antibody (Product # PA5-30581) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Enrichment of endogenous DNMT1 protein at specific gene loci using Anti-DNMT1 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-DNMT1 Rabbit Polyclonal Antibody (Product # PA5-30581, 4 ug) on sheared chromatin from 2 million HeLa cells using the MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over mir-137 and POMC (positive) and SAT2 satellite repeats (negative). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Cross-linked ChIP was performed with HCT116 chromatin extract and 5 µg of either control rabbit IgG or DNMT1 Polyclonal Antibody (Product # PA5-30581). The precipitated DNA was detected by PCR with primer set targeting to ILIR2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Enrichment of endogenous DNMT1 protein at specific gene loci using Anti-DNMT1 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-DNMT1 Rabbit Polyclonal Antibody (Product # PA5-30581, 4 ug) on sheared chromatin from 2 million HeLa cells using the MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over mir-137 and POMC (positive) and SAT2 satellite repeats (negative). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Cross-linked ChIP was performed with HCT116 chromatin extract and 5 µg of either control rabbit IgG or DNMT1 Polyclonal Antibody (Product # PA5-30581). The precipitated DNA was detected by PCR with primer set targeting to ILIR2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: DNMT1 Polyclonal Antibody immunoprecipitates DNMT1 protein in IP experiments. IP samples: HeLa nuclear extract. A. Control with 4 µg of preimmune Rabbit IgG. B. Immunoprecipitation of DNMT1 protein by 4 µg DNMT1 Polyclonal Antibody (Product # PA5-30581). 5 % SDS-PAGE. The immunoprecipitated DNMT1 protein was detected by DNMT1 Polyclonal Antibody (Product # PA5-30581) diluted at 1:1,000.

PA5-30581

Antibody data