Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Dnmt3a with ab188470 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab188470 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue sections labeling Dnmt3a with purified ab188470 at 1/2000 (0.409 μg/ml). Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Dnmt3a with purified ab188470 at 1/80 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling Dnmt3a with purified ab188470 at 1/2000 (0.409 μg/ml). Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling Dnmt3a with purified ab188470 at 1/2000 (0.409 μg/ml). Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470).

• WB

Supplier Data

Western blot - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

This data was developed using the same antibody clone in a different buffer formulation (ab188470).

Blocking/Dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (J Biol Chem. 2002. 277, 38746-38754. PMID : 12138111).

All lanes:

Western blot - Anti-Dnmt3a antibody [EPR18455] (<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr18455-ab188470'>ab188470</a>) at 1/10000 dilution

All lanes:

HEK-293 (Human epithelial cells from embryonic kidney) 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 1/50000 dilution

Predicted band size: 102 kDa,83 kDa

Observed band size: 130 kDa,74 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

Lanes 1 - 4 : Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab188470 was shown to recognize Dnmt3a when Dnmt3a knockout samples were used, along with additional cross-reactive bands. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and ab8245 (loading control to GAPDH) were diluted to 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470).

All lanes:

Western blot - Anti-Dnmt3a antibody [EPR18455] (<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr18455-ab188470'>ab188470</a>) at 1/5000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

Dnmt3a knockout HAP1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Predicted band size: 102 kDa

false

• WB

Lab

Western blot - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

This data was developed using the same antibody clone in a different buffer formulation (ab188470).

Lanes 1-4 : Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.

ab188470 Anti-Dnmt3a antibody [EPR18455] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Dnmt3a antibody [EPR18455] (<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr18455-ab188470'>ab188470</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Dnmt3a knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human DNMT3A knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-dnmt3a-knockout-hela-cell-line-ab261793'>ab261793</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 102 kDa

Observed band size: 125 kDa

false

• WB

Supplier Data

Western blot - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

This data was developed using the same antibody clone in a different buffer formulation (ab188470).

Blocking/Dilution buffer : 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (J Biol Chem. 2002. 277, 38746-38754. PMID : 12138111).

All lanes:

Western blot - Anti-Dnmt3a antibody [EPR18455] (<a href='/en-us/products/primary-antibodies/dnmt3a-antibody-epr18455-ab188470'>ab188470</a>) at 1/10000 dilution

All lanes:

HeLa (Human epithelial cells from cervix adenocarcinoma) 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 1/50000 dilution

Predicted band size: 102 kDa

Observed band size: 130 kDa

false

Exposure time: 5s

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-Dnmt3a antibody [EPR18455] - BSA and Azide free (AB232391)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188470)

CUT&RUN profiling with Dnmt3a antibody reveals the expected genomic enrichment pattern in cells. Representative genome browser tracks show CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with Dnmt3a antibody (Abcam ab188470, 0.5 µg). 500,000 HeLa cells were used per reaction. IgG, H3K4me3, and H3K27me3 antibodies were included as controls to assess non-specific background, active promoters, and repressed chromatin, respectively. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Images were generated using Integrative Genomics Viewer (IGV, Broad Institute).