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Immunocytochemistry analysis of SK-Br-3 cells labeling Cytokeratin 7 with Rabbit anti-Cytokeratin 7 antibody (HA720144F) 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 7 antibody (HA720144F, 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.
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Immunofluorescence analysis of paraffin-embedded human liver tissue labeling Cytokeratin 7 (HA720144F) and Cytokeratin 8 (HA720132F).
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 7 (HA720144F, red) at 1/200 dilution and Cytokeratin 8 (HA720132F, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.
DAPI was used as nuclear counterstain.
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Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 7 (HA720144F), Cytokeratin 8 (HA720132F) 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 7 (HA720144F, red) at 1/50 dilution, Cytokeratin 8 (HA720132F, green) at 1/200 dilution and Vimentin (EM0401, yellow) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.
Alexa Fluor® 555 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. DAPI was used as nuclear counterstain.
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Immunofluorescence analysis of paraffin-embedded human breast tissue labeling Cytokeratin 7 (HA720144F).
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 antibody Cytokeratin 7 (HA720144F, iFluor™ 647) at 1/200 dilution overnight at 4 ℃, washed with PBS. DAPI was used as nuclear counterstain.
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Flow cytometric analysis of SK-Br-3 cells labeling Cytokeratin 7.
Cells were fixed and permeabilized. Then incubated for 1 hour at +4℃ with Cytokeratin 7 (HA720144F, red, 1ug/ml). Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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