Cytokeratins comprise a diverse group of intermediate filament proteins (IFPs) that are expressed as pairs in both keratinized and non-keratinized epithelial tissue, where they constitute up to 85% of mature keratinocytes in the vertebrate epidermis. Cytokeratins play a critical role in differentiation and tissue specialization and function to maintain the overall structural integrity of epithelial cells. The a-helical coiled-coil dimers associate laterally end-to-end to form 10 nm diameter filaments. Cytokeratins are useful markers of tissue differentiation and, in addition, they aid in the characterization of malignant tumors. Cytokeratin 7 (also known as sarcolectin) agglutinates normal and transformed cells with a high affinity for simple sugars. Cytokeratin 7 also inhibits the synthesis of interferon-dependent secondary proteins thus reversing the antiviral effect of interferon induction and restoring cells to their status ad primum. In normal and transformed cells, Cytokeratin 7 localizes to the membrane.
Background References
1. Hrudka J. et al. Cytokeratin 7 expression as a predictor of an unfavorable prognosis in colorectal carcinoma. Sci Rep. 2021 Sep
2. Statz E. et al. Cytokeratin 7, GATA3, and SOX-10 is a Comprehensive Panel in Diagnosing Triple Negative Breast Cancer Brain Metastases. Int J Surg Pathol. 2021 Aug
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.
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.
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.
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.
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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