Monospecific monoclonal antibodies to keratin 1 carboxy terminal (synthetic peptide) and to keratin 10 as markers of epidermal differentiation

Abstract

Monospecific antibodies to individual keratin polypeptides can be used to examine the tissue and cellular coexpression of members of keratin pairs. Monospecific monoclonal and polyclonal antibodies have been raised to keratins 1 and 10 using both crude cytoskeletal extracts and synthetic peptides. The tissue distribution of these keratins has been determined against a panel of freshly frozen normal tissues from humans, rodents and pigs. Epidermal expression has been examined in psoriatic plaques, and healing wounds, as examples of epidermal hyperproliferation. Cultured keratinocytes in monolayer (low calcium), stratified (high calcium), and complex cultures, transformed keratinocytes, and tumour cell lines, have been examined for the in vitro expression of these keratins. The sensitivity and precise localization of reactivity with these monospecific antibodies gives a highly accurate picture of individual cell expression. There is confirmation of coexpression of keratins 1 and 10 in epidermal and mucosal sites, and with keratin 16 in hyperproliferative states. These monospecific antibodies provide an important means of examining keratin expression in epidermal tumours and keratinizing disorders, and of seeking keratin mutations in cell lines and in skin diseases.

K15 Expression Implies Lateral Differentiation within Stratified Epithelial Basal Cells

“…Other keratin antibodies were LE41 (K8) and LE61 (K18) (Lane, 1982), LL002 (K14) (Purkis et al, 1990), LL025 (K16) (Wilson et al, 1994), LHK6 (K6) and LHK15 (K15) (Waseem et al, 1999), LP2K (K19) (Stasiak et al, 1989) RCK102 (K5 &K8) (Broers et al, 1986), and LH2 (K10) (Leigh et al, 1993). Additional monoclonal antibodies used were E-cadherin (Transduction Laboratories, Lexington, Kentucky) diluted 1:50, 11-5F (anti-desmoplakin) (Parrish et al, 1987) diluted 1:100; and MM-1 (anti-Ki67) (Novacastra, Newcastle upon Tyne, United Kingdom).…”

A functional "knockout" of human keratin 14.

“…), and LHPl to KIO (Leigh et al 1993). A polyclonal antiserum to the carboxy-terminal peptide of K5 (RbaK5; Purkis et al 1990) and another to the carboxy-terminal peptide of Kl (RbaKl; Leigh et al 1993) were also used. Second antibodies used were rabbit anti-mouse immunoglobulins and swine anti-rabbit immunoglobulins conjugated with FITC (DAKO), diluted 1:200 in PBS/1% BSA for tissue staining, and sheep anti-mouse immunoglobulin conjugated to FITC (Sigma), diluted 1:50 in expired tissue culture medium/10% fetal calf serum, for cell staining.…”

“…comm. ), and LHPl to KIO (Leigh et al 1993). A polyclonal antiserum to the carboxy-terminal peptide of K5 (RbaK5; Purkis et al 1990) and another to the carboxy-terminal peptide of Kl (RbaKl; Leigh et al 1993) were also used.…”

Addition of Selenium to <i>Carica papaya</i> Linn Pulp Extract Enhances Dermal Wound Healing Activity

“…Fibroblasts and keratinocytes require optimal calcium signalling for cellular migration and differentiation leading to re-epithelization during the proliferative phase of wound healing [14,15]. Se, the most abundant micronutrient in C. papaya, is an essential cofactor for endogenous antioxidant enzymes which can reduce oxidative tissue damage resulting from neutrophil infiltration and microbial invasion of wounds [16].…”