- GTX22851 - GeneTex DNMT3B antibody

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Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Western blot analysis of DNA Methyltransferase 3 was performed by loading 25 ug of Hela (Lane 1), A549 (Lane 2), and NIH-3T3 (Lane 3) cell lysates and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4?C overnight. The membrane was probed with a DNA Methyltransferase 3 polyclonal antibody (GTX22851) at a dilution of 1:1000 overnight at 4¢XC, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Results show a band at 97 kDa in A549 and Hela cell lysates.

Supportive validation

Submitted by: GeneTex (provider)
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Experimental details: Immunohistochemistry analysis of DNA Methyltransferase 3 showing positive staining in the nucleus and cytoplasm of paraffin-treated Human breast carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a DNA Methyltransferase 3 polyclonal antibody (GTX22851) diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4¢XC in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Immunohistochemistry analysis of DNA Methyltransferase 3 showing positive staining in the nucleus of paraffin-treated Human testis tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a DNA Methyltransferase 3 polyclonal antibody (GTX22851) diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4¢XC in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: GeneTex (provider)
Main image
Experimental details: Immunohistochemistry analysis of DNA Methyltransferase 3 showing positive staining in the nucleus and cytoplasm of paraffin-treated Mouse brain tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a DNA Methyltransferase 3 polyclonal antibody (GTX22851) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4¢XC in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

GTX22851