All lanes : Anti-STIP1/STI1 antibody [EPR6605] (ab126724) at 1/10000 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : STIP1 knockout HAP1 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HEK-293 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 63 kDa
Observed band size: 63 kDa

This data was developed using the same antibody clone in a different buffer formulation (ab126724).

Lanes 1 - 4: Merged signal (red and green). Green - ab126724 observed at 63 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab126724 was shown to react with STIP1/STI1 in wild-type HAP1 cells in western blot with loss of signal observed in STIP1 knockout sample. Wild-type and STIP1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab126724 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

This image was made using ab126724 which is the same antibody as ab238963 with BSA and Azide
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human thyroid cancer tissue sections labeling STIP1/STI1 with Purified ab126724 at 1:1000 dilution (0.18 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling STIP1/STI1 (red) with ab126724 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126724).

This image was made using ab126724 which is the same antibody as ab238963 with BSA and Azide
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human testis tissue sections labeling STIP1/STI1 with Purified ab126724 at 1:1000 dilution (0.18 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Unpurified ab126724, at a dilution of 1/250, staining STIP1/STI1 in paraffin-embedded human ovarian carcinoma tissue by Immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, tissue culture supernatant and sodium azide (ab126724).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.