iFluor™ 647 Conjugated Cytokeratin 8 Recombinant Rabbit Monoclonal Antibody [SU0338]

Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 8 (HA720145F) and Vimentin (EM0401).

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS. And then probed with the primary antibodies Cytokeratin 8 (HA720145F, red) at 1/200 dilution and Vimentin (EM0401, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.

iFluor™ 488 conjugate-Goat anti-Mouse IgG (HA1125) was used as the secondary antibody at 1/1,000 dilution. DAPI was used as nuclear counterstain.

Immunocytochemistry analysis of SK-Br-3 cells labeling Cytokeratin 8 with Rabbit anti-Cytokeratin 8 antibody (HA720145F) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% normal goat serum for 1 hour at 37℃. Cells were then incubated with Rabbit anti-Cytokeratin 8 antibody (HA720145F, red) at 1/100 dilution in 2% normal goat serum overnight at 4 ℃. Nuclear DNA was labelled in blue with DAPI.

Beta III Tubulin (M0805-8, green) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/800 dilution.

Flow cytometric analysis of SK-Br-3 cells labeling Cytokeratin 8 (HA720145F, iFluor™ 647).

Cells were fixed and permeabilized. Then incubated for 1 hour at +4℃ with Cytokeratin 8 (HA720145F, red, 1/100) and Rabbit IgG Isotype Control (iFluor™ 647, green, 1/100). Unlabelled sample was used as a control (cells without incubation with primary antibody; black).