Recombinant Anti-Cytokeratin 20 antibody [SP33] (ab64090)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling Cytokeratin 20 with ab64090 at 1/100 dilution (17.9 µg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on rat colon, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64090 for 30 mins at room temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue sections labeling Cytokeratin 20 with ab64090 at 1/100 dilution (17.9 µg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on human colon carcinoma, performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64090 for 30 mins at room temperature.

Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for Cytokeratin 20 with ab64090 at a 1/100 dilution in immunohistochemical analysis.

Immunohistochemistry (Frozen) analysis of rat colon tissue section labeling Cytokeratin 20 with purified ab64090 at 1/1000 (1.8 µg/ml). Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunocytochemistry/ Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma epithelial cell) cells labeling Cytokeratin 20 with purified ab64090 at 1/500 (3.58 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 was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow cytometry analysis of HT-29 (human colorectal adenocarcinoma) labeling Cytokeratin 20 with purified ab64090 at 1/1700 dilution (1.05 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control -Rabbit monoclonal IgG (ab172730) (Black). Unlableled control -Unlabelled cells (blue).