Anti-KLF4 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KLF4 antibody (AB129473)

ab129473 staining KLF4 in mES cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab129473 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• WB

Lab

Western blot - Anti-KLF4 antibody (AB129473)

Gel type : MOPS

Blocking buffer : 2% BSA block

All lanes:

Western blot - Anti-KLF4 antibody (ab129473) at 1 µg/mL

Lane 1:

Mouse embryonic stem cell lysate at 20 µg

Lane 2:

F9 whole cell lysate at 20 µg

Lane 3:

LoVo whole cell lysate at 20 µg

Lane 4:

HEK-293 whole cell lysate at 20 µg

Lane 5:

HT29 whole cell lysate at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 54 kDa

Observed band size: 60 kDa

false

Exposure time: 1min

• WB

Unknown

Western blot - Anti-KLF4 antibody (AB129473)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab129473 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-KLF4 antibody (ab129473) at 1 µg/mL

All lanes:

E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate at 10 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 1/10000 dilution

Predicted band size: 54 kDa

Observed band size: 60 kDa

true

Exposure time: 8min