Anti-Dnmt3L antibody [EPR18774]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Immunohistochemical analysis of paraffin-embedded Human yolk sac tumor tissue labeling Dnmt3L with ab194094 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nuclear staining was found on the tumor cells of Human yolk sac tumor.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized NCCIT (Human pluripotent embryonic carcinoma) and HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Dnmt3L with ab194094 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on NCCIT cell line. Negative expression in HeLa cells (PMID : 20460473).

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab194094 at 1/250 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Immunohistochemical analysis of paraffin-embedded Human seminoma tissue labeling Dnmt3L with ab194094 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Mainly nuclear with weak cytoplasmic staining was found on tumor cells of Human seminoma.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Dnmt3L with ab194094 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on the Human liver is observed.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Dnmt3L with ab194094 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on the Human liver is observed.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Blocking/Dilution buffer : 5% NFDM/TBST.

Observed MW : 48 kDa (mouse), 44 kDa (human).

Exposure time : Lanes 1 & 2 : 1 second; Lanes 3,4,5,6 & 7 : 3 minutes. GAPDH control lanes : 30 seconds.

1. The predicted molecular weight of human Dnmt3L is 44kDa and the predicted molecular weight of mouse Dnmt3L is 48kDa. This is consistent with what has been observed.
2. HeLa and T-47D cells do not express Dnmt3L (PMID : 20460473).

All lanes:

Western blot - Anti-Dnmt3L antibody [EPR18774] (ab194094) at 1/1000 dilution

Lane 1:

mESC (Mouse embryonic stem cells ) whole cell lysate at 20 µg

Lane 2:

F9 (Mouse embryonic testicular cancer cell line) whole cell lysate at 20 µg

Lane 3:

Rat testis lysate at 20 µg

Lane 4:

NTERA-2 cl.D1 [NT2/D1] (Human malignant pluripotent embryonic carcinoma) whole cell lysate at 20 µg

Lane 5:

NCCIT (Human pluripotent embryonic carcinoma) whole cell lysate at 20 µg

Lane 6:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 7:

T-47D (Human ductal breast epithelial tumor cell line) whole 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 1/100000 dilution

Predicted band size: 44 kDa

Observed band size: 44 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Dnmt3L antibody [EPR18774] (AB194094)

Dnmt3L was immunoprecipitated from 1mg of F9 (Mouse embryonic testicular cancer cell line) whole cell lysate with ab194094 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab194094 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : F9 whole cell lysate,10ug (Input).

Lane 2 : ab194094 IP in F9 whole cell lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab191594 in F9 whole cell lysate.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Dnmt3L antibody [EPR18774] (ab194094)

Predicted band size: 44 kDa

Observed band size: 48 kDa

false