Anti-STELLAR antibody

• IHC-Fr

Characteriser

Immunohistochemistry (Frozen sections) - Anti-STELLAR antibody (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.

This image is courtesy of Petra Hajkova, University of Cambridge

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STELLAR antibody (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 +4°C. 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-STELLAR antibody (AB19878)

ab19878 staining STELLAR 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 ab19878 at 5µg/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 magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-STELLAR antibody (AB19878)

Immunocytochemistry-immunofluorescence using Anti-STELLAR antibody, ab19878. Publication image from Kim, Y. et al., 2020, Nat Commun, 32332736. Legend direct from paper.

PGCs show rapid non-canonical Hh signalling responses.a Immunofluorescence of Smo, p-Creb, Gli3 and p-Src, co-stained with germ cell marker Stella in GR primary culture cells with or without Shh treatment for 10 min. Representative images from at least three biologically independent experiments are shown. Scale bar, 10 µm. b The relative fluorescence intensity values of Smo, p-Creb, Gli3 and p-Src, normalised to the signal intensity of Stella observed in PGCs were compared with or without Shh treatment. Unpaired t-test with Welch’s correction indicates a significant increase in Smo (DMF (n = 14), Shh (n = 13), ****P < 0.0001), p-Creb (DMF (n = 9), Shh (n = 7), ****P < 0.0001) and p-Src (DMF (n = 10), Shh (n = 11), ****P < 0.0001) but no significant change in Gli3 (DMF (n = 9), Shh (n = 9), P = 0.8542). Error bars represent SD. Unpaired t-test with Welch’s correction. Source data are provided as a Source Data file.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-STELLAR antibody (AB19878)

Immunocytochemistry-immunofluorescence using Anti-STELLAR antibody, ab19878. Publication image from Kim, Y. et al., 2020, Nat Commun, 32332736. Legend direct from paper.

PGCs are naturally unciliated but still responsive to Hh signalling.a Primary cultures of GR were treated with DMF or Pur for 18 h and analysed by immunofluorescence staining for Smo and Gli3. The positive and negative staining for Stella, a germ cell marker, distinguished PGCs and somatic cells, respectively. The merged images are shown without Stella signal for improved clarity. Representative images from three biologically independent experiments are shown. Scale bar, 10 µm. b Dot plots showing the relative fluorescence intensity values of Smo and Gli3 signal observed in PGCs treated with DMF or Pur, which were normalised to the fluorescence intensity values of Stella in each cell. Unpaired t-test with Welch’s correction indicates a significant increase in Smo (DMF (n = 9), Pur (n = 31), P < 0.0001) and Gli3 (DMF (n = 18), Pur (n = 20), P < 0.0001). c Cilia staining of primary cultures of dissected E10.5 mouse GR tissues. Arl13b and acetylated tubulin are used for axoneme staining and gamma-tubulin and CEP164 are used for basal body staining. SSEA1 and Stella are used as germ cell markers. Representative images from three independent experiments are shown. Scale bar, 10 µm. d Ciliation frequency of the PGCs (n = 158) and the somatic cells (n = 971) observed from GR cultures. Error bars represent SD of three independent experiments. Each data point represents one experiment. Unpaired t-test, two-tailed (****P < 0.0001). Source data are provided as a Source Data file.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-STELLAR antibody (AB19878)

Immunocytochemistry-immunofluorescence using Anti-STELLAR antibody, ab19878. Publication image from Kim, Y. et al., 2020, Nat Commun, 32332736. Legend direct from paper.

PGCs are naturally unciliated but still responsive to Hh signalling.a Primary cultures of GR were treated with DMF or Pur for 18 h and analysed by immunofluorescence staining for Smo and Gli3. The positive and negative staining for Stella, a germ cell marker, distinguished PGCs and somatic cells, respectively. The merged images are shown without Stella signal for improved clarity. Representative images from three biologically independent experiments are shown. Scale bar, 10 µm. b Dot plots showing the relative fluorescence intensity values of Smo and Gli3 signal observed in PGCs treated with DMF or Pur, which were normalised to the fluorescence intensity values of Stella in each cell. Unpaired t-test with Welch’s correction indicates a significant increase in Smo (DMF (n = 9), Pur (n = 31), P < 0.0001) and Gli3 (DMF (n = 18), Pur (n = 20), P < 0.0001). c Cilia staining of primary cultures of dissected E10.5 mouse GR tissues. Arl13b and acetylated tubulin are used for axoneme staining and gamma-tubulin and CEP164 are used for basal body staining. SSEA1 and Stella are used as germ cell markers. Representative images from three independent experiments are shown. Scale bar, 10 µm. d Ciliation frequency of the PGCs (n = 158) and the somatic cells (n = 971) observed from GR cultures. Error bars represent SD of three independent experiments. Each data point represents one experiment. Unpaired t-test, two-tailed (****P < 0.0001). Source data are provided as a Source Data file.