Anti-Lefty antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lefty antibody (AB22569)

ab22569 staining Lefty in mES cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 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 ab22569 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).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

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

• WB

Unknown

Western blot - Anti-Lefty antibody (AB22569)

The mouse recombinant Lefty protein consists of two bands of ~34 and 12 kDa. Of these, ab22569 recognises the 34 kDa band. The human recombinant Lefty protein consists of three bands of 44, 38 and 34 kDa. Of these, the first two are recognised by ab22569. The protein sizes differ from the expected molecular weight of Lefty due to proteolytic processing and glycosylation.

All lanes:

Western blot - Anti-Lefty antibody (ab22569) at 1 µg/mL

Lane 1:

Mouse Lefty1 Recombinant Protein at 0.01 µg

Lane 2:

Human LeftyA Recombinant Protein at 0.01 µg

Secondary

All lanes:

Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution

Predicted band size: 41 kDa

Observed band size: 34 kDa,38 kDa,44 kDa

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