Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat spleen tissue sections labeling RPS6 with ab101691 at 1/100 dilution (1.23 µg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Dako™REAL™ EnVision™ Detection System (K5007) was used as the secondary antibody. Hematoxylin was used as a counterstain. Cytoplasmic staining on human rat spleen without alkaline phosphatase treatment (image A); and no signal was detected when tissues were treated with alkaline phosphatase (image B), performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab101691 for 10 mins at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse stomach tissue sections labeling RPS6 with ab101691 at 1/100 dilution (1.23 µg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Dako™REAL™ EnVision™ Detection System (K5007) was used as the secondary antibody. Hematoxylin was used as a counterstain. Cytoplasmic staining on mouse stomach without alkaline phosphatase treatment (image A); and no signal was detected when tissues were treated with alkaline phosphatase (image B), performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab101691 for 10 mins at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling RPS6 with ab101691 at 1/100 dilution (1.23 µg/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Dako™ REAL™ EnVision™ Detection System (K5007) was used as the secondary antibody. Hematoxylin was used as a counterstain. Cytoplasmic staining on human breast carcinoma without alkaline phosphatase treatment (image A); and no signal was detected when tissues were treated with alkaline phosphatase (image B), performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab101691 for 10 mins at room temperature.

Immunocytochemistry/ Immunofluorescence analysis of PC-3 (human prostate adenocarcinoma epithelial cell) treated with Lamda phosphatase cells labeling RPS6 with purified ab101691 at 1:500 (0.25 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Dot blot analysis of RPS6 (phospho S235 + S236) peptide (Lane 1), RPS6 (phospho S235) peptide (phospho 2) RPS6 (phospho S236) peptide (Lane 3) and RPS6 non-phospho peptide (Lane 4) labeling RPS6 (phospho S235 + S236) peptide with purified ab101691 at 0.123 ug/mL. ab97051 was used as the secondary antibody at 0.01 ug/mL.

Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST .

Flow Cytometry analysis of PC-3 (Human prostate adenocarcinoma epithelial cell) cells labeling RPS6 with purified ab101691 at 1:200 dilution (0.615 µg/ml) Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / Blue.

ab101691 at 1/100 dilution staining S6K in Human breast carcinoma by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.