ab133350 staining CD90 / Thy1  in the SH-SY5Y human neuroblastoma epithelial cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/100). ab150077 an Alexa Fluor® 488-conjugated Goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI was used as a nuclear counter stain and Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counter stain microtubules.

All lanes : Anti-c-Kit antibody [YR145] (ab32363) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate

Lane 2 : KIT knockout HAP1 whole cell lysate

Lane 3 : HUVEC whole cell lysate

Lysates/proteins at 40 µg per lane.

Predicted band size: 110 kDaLanes 1 - 3: Merged signal (red and green). Green - ab32363 observed at 109 kDa. Red - loading control, ab9484, observed at 37 kDa.

Lanes 1 - 3: Merged signal (red and green). Green - ab32363 observed at 109 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab32363 was shown to recognize 0 in wild-type HAP1 cells as signal was lost at the expected MW in KIT knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and KIT knockout samples were subjected to SDS-PAGE. Ab32363 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human seminoma tissue labelling c-Kit with unpurified ab32363.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD38 with purified ab108403 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Immunohistochemical analysis of paraffin-embedded Human normal kidney tissue labelling CD90 / Thy1 with ab133350 at 1/100 dilution.

Immunohistochemical analysis of CD59 in paraffin embedded Human placenta tissue labelled with ab133707 at a 1/100 dilution.

Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
Lane 2: CD59 knockout HAP1 whole cell lysate (40 µg)

Lanes 1 - 2: Merged signal (red and green). Green - Anti-CD59 antibody [EPR6425(2)] (ab133707) observed at 14 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab133707 was shown to specifically react with CD59 in wild-type HAP1 cells as signal was lost in CD59 knockout cells. Wild-type and CD59 knockout samples were subjected to SDS-PAGE. Ab133707 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human angiosarcoma labeling CD34 with unpurified ab81289 at 1/100-1/250.

Immunocytochemistry/Immunofluorescence analysis of HUVEC (Human umbilical vein endothelial cell line) cells labelling CD34 with purified ab81289 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain.

Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 488-conjugated goat anti-mouse IgG (1/500).