Anti-Stella antibody - Primordial Germ Cell Marker (ab19878)

Immunohistochemistry (Frozen sections) - Stella antibody - Germ Cell Marker (ab19878)

The image shows ab19878 staining of a crysosection of mouse embryonic genital ridges (E14.5). The samples were fixed overnight in 4% paraformaldehyde, permeabilised with 0.1% Triton and stained overnight at 4 degrees. 4µg/ml of antibody was used. Staining was in the nucleus and cytoplasm of Oct4-positive cells. The blue fluorescence is DAPI staining of DNA.

Petra Hajkova, University of Cambridge

Western blot - Anti-Stella antibody - Primordial Germ Cell Marker (ab19878)

All lanes : Anti-Stella antibody - Primordial Germ Cell Marker (ab19878) at 1 µg/ml

Lane 1 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate
Lane 2 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate
Lane 3 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate
Lane 4 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate with Stella peptide (ab23324) at 2 µg/ml
Lane 5 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate with Stella peptide (ab23324) at 2 µg/ml
Lane 6 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate with Stella peptide (ab23324) at 2 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size : 17 kDa
Observed band size : 23 kDa (why is the actual band size different from the predicted?)
Additional bands at : 125 kDa,25 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 3 minutes

Immunocytochemistry/ Immunofluorescence - Anti-Stella antibody - Primordial Germ Cell Marker (ab19878)

ICC/IF image of ab19878 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19878, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunocytochemistry/ Immunofluorescence-Anti-Stella antibody - Primordial Germ Cell Marker(ab19878)

ICC/IF image of ab19878 stained mouse embryonic stem cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19878, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.