Cytokeratins comprise a diverse group of intermediate filament proteins that are expressed as pairs in both keratinized and non-keratinized epithelial tissue. The cytokeratin proteins play a critical role in differentiation, as well as tissue specialization and function, to maintain the overall structural integrity of epithelial cells. Cytokeratins are also useful markers in identifying the origin of metastatic tumors. Cytokeratin 16 is expressed in benign stratified squamous epithelium and squamous cell carcinoma of the head and neck, as well as luminal cells of mammary gland and sweat ducts. It is absent in noninvasive breast carcinomas and normal breast tissue. Mutations in the Cytokeratin 16 gene cause various diseases, including pachyonychia congenita type 1 (PC1), nonepidermolytic palmoplantar keratoderma (NEPPK) and unilateral palmoplantar verrucous nevus (UPVN).
Background References
1. Pal SK. et. al. The expression profiles of acidic epithelial keratins in ameloblastoma. Oral Surg Oral Med Oral Pathol Oral Radiol 115:523-31 (2013).
2. Cheng CH. et. al. Keratin gene expression profiles after digit amputation in C57BL/6 vs. regenerative MRL mice imply an early regenerative keratinocyte activated-like state. Physiol Genomics 45:409-21 (2013).
Immunofluorescence analysis of paraffin-embedded human esophagus tissue labeling Cytokeratin 16 (HA720137F) 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 16 (HA720137F, green) at 1/50 dilution and Vimentin (EM0401, red) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.
iFluor™ 594 conjugate-Goat anti-Mouse IgG (HA1126) was used as the secondary antibody at 1/1,000 dilution. DAPI was used as nuclear counterstain.
Immunofluorescence analysis of paraffin-embedded human skin tissue labeling Cytokeratin 16 (HA720137F) 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 16 (HA720137F, green) at 1/200 dilution and Vimentin (EM0401, red) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.
iFluor™ 594 conjugate-Goat anti-Mouse IgG (HA1126) was used as the secondary antibody at 1/1,000 dilution. DAPI was used as nuclear counterstain.
Immunocytochemistry analysis of HepG2 cells labeling Cytokeratin 16 with Rabbit anti-Cytokeratin 16 antibody (HA720137F) at 1/100 dilution.
Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Cytokeratin 16 antibody (HA720137F) at 1/100 dilution in 1% BSA overnight at 4 ℃. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) were used as the secondary antibody at 1/800 dilution.
Immunocytochemistry analysis of A431 cells labeling Cytokeratin 16 with Rabbit anti-Cytokeratin 16 antibody (HA720137F) at 1/50 dilution.
Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Cytokeratin 16 antibody (HA720137F) at 1/50 dilution in 1% BSA overnight at 4 ℃. Nuclear DNA was labelled in blue with DAPI.
Flow cytometric analysis of A431 cells labeling Cytokeratin 16.
Cells were fixed and permeabilized. Then incubated for 1 hour at +4℃ with Cytokeratin 16 (HA720137F, red, 1ug/ml). Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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