Anti-WTAP antibody [EPR18744] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling WTAP with ab195380 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Jurkat cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody  at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab195380 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody 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 (ab195380).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling WTAP with ab195380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human endometrium is observed. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195380).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling WTAP with ab195380 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab195380 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody 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 (ab195380).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling WTAP with ab195380 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195380).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling WTAP with ab195380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human liver cancer is observed. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195380).

• IP

Supplier Data

Immunoprecipitation - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

WTAP was immunoprecipitated from 0.35 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with ab195380 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab195380 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Jurkat whole cell lysate 10μg (Input).

Lane 2 : ab195380 IP in Jurkat whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab195380 in Jurkat whole cell lysate.

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

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195380).

All lanes:

Immunoprecipitation - Anti-WTAP antibody [EPR18744] (<a href='/en-us/products/primary-antibodies/wtap-antibody-epr18744-ab195380'>ab195380</a>)

Predicted band size: 44 kDa

false

• WB

Supplier Data

Western blot - Anti-WTAP antibody [EPR18744] - BSA and Azide free (AB232392)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : WTAP knockout HAP1 whole cell lysate (20 μg)
Lane 3 : MOLT-4 whole cell lysate (20 μg)
Lane 4 : K562 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab195380 observed at 50 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab195380 was shown to specifically react with WTAP in wild-type HAP1 cells as signal was lost in WTAP knockout cells. Wild-type and WTAP knockout samples were subjected to SDS-PAGE. ab195380 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed 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 (ab195380).

All lanes:

Western blot - Anti-WTAP antibody [EPR18744] (<a href='/en-us/products/primary-antibodies/wtap-antibody-epr18744-ab195380'>ab195380</a>)

Predicted band size: 44 kDa

false