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The monoclonal antibody EPR1614Y against the stem cell biomarker keratin K15 lacks specificity and reacts with other keratins
- Source :
- Scientific Reports, Vol 9, Iss 1, Pp 1-12 (2019), Scientific Reports
- Publication Year :
- 2019
- Publisher :
- Nature Publishing Group, 2019.
-
Abstract
- Keratin 15 (K15), a type I keratin, which pairs with K5 in epidermis, has been used extensively as a biomarker for stem cells. Two commercial antibodies, LHK15, a mouse monoclonal and EPR1614Y, a rabbit monoclonal, have been widely employed to study K15 expression. Here we report differential reactivity of these antibodies on epithelial cells and tissue sections. Although the two antibodies specifically recognised K15 on western blot, they reacted differently on skin sections and cell lines. LHK15 reacted in patches, whereas EPR1614Y reacted homogenously with the basal keratinocytes in skin sections. In cultured cells, LHK15 did not react with K15 deficient NEB-1, KEB-11, MCF-7 and SW13 cells expressing only exogenous K8 and K18 but reacted when these cells were transduced with K15. On the other hand, EPR1614Y reacted with these cells even though they were devoid of K15. Taken together these results suggest that EPR1614Y recognises a conformational epitope on keratin filaments which can be reconstituted by other keratins as well as by K15. In conclusion, this report highlights that all commercially available antibodies may not be equally specific in identifying the K15 positive stem cell.
- Subjects :
- 0301 basic medicine
Keratinocytes
medicine.drug_class
Blotting, Western
Intermediate Filaments
lcsh:Medicine
Cross Reactions
Monoclonal antibody
Sensitivity and Specificity
Article
Cell Line
03 medical and health sciences
0302 clinical medicine
Antibody Specificity
Keratin
medicine
Animals
Humans
lcsh:Science
Cells, Cultured
Skin
chemistry.chemical_classification
Multidisciplinary
Epidermis (botany)
biology
integumentary system
Chemistry
Keratin-15
Stem Cells
lcsh:R
Antibodies, Monoclonal
Epithelial Cells
Molecular biology
Blot
Cytoskeletal Proteins
030104 developmental biology
Epidermal Cells
Cell culture
Monoclonal
biology.protein
Keratins
lcsh:Q
Stem cell
Antibody
Epidermis
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 20452322
- Volume :
- 9
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....085e1241e302a5bfb9a30f6ca49f0dae