Anti-WSTF antibody [EP1704Y] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This data was developed using ab51256, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling WSTF with Purified ab51256 at 1 : 20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This data was developed using ab51256, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling WSTF with Purified ab51256 at 1 : 700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This data was developed using ab51256, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling WSTF with Purified ab51256 at 1 : 50 dilution (2.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This data was developed using ab51256, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue sections labeling WSTF with Purified ab51256 at 1 : 700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This data was developed using ab51256, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cardiac muscle tissue sections labeling WSTF with Purified ab51256 at 1 : 700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• WB

Unknown

Western blot - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This WB data was generated using the same anti-WSTF antibody clone, EP1704Y, in a different buffer formulation (cat# ab51256).

Lanes 1 - 4 : Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab51265 was shown to recognize WSTF in wilt-type cells along with additional cross-reactive bands as signal was lost in WSTF knockout samples. Wild-type and WSTF knockout samples were subjected to SDS-PAGE. ab51256 and ab18058 (loading control to vinculin) were diluted 1/15 000 and 1/10 000 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.

All lanes:

Western blot - Anti-WSTF antibody [EP1704Y] (<a href='/ko/products/primary-antibodies/wstf-antibody-ep1704y-ab51256'>ab51256</a>) at 1/15000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

WSTF knockout HAP1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

PC12 cell lysate at 20 µg

Predicted band size: 171 kDa

false

• WB

Lab

Western blot - Anti-WSTF antibody [EP1704Y] - BSA and Azide free (AB235388)

This data was developed using the same antibody clone in a different buffer formulation (ab51256).

Lanes 1- 2 : Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab51256 was shown to react with WSTF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264907 (knockout cell lysate ab257370) was used. Wild-type HeLa and BAZ1B knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51256 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 15000 dilution and a 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-WSTF antibody [EP1704Y] (<a href='/ko/products/primary-antibodies/wstf-antibody-ep1704y-ab51256'>ab51256</a>) at 1/15000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

BAZ1B knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human BAZ1B (WSTF) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-baz1b-wstf-knockout-hela-cell-line-ab264907'>ab264907</a>)

Predicted band size: 171 kDa

Observed band size: 175 kDa

false