A unique monoclonal antibody 29A stains the cytoplasm of amniotic epithelia and cutaneous basement membrane.
BACKGROUND
The basic function of epithelia is to provide a boundary between tissue and its external environment, and is achieved by a wide variety of components including extracellular molecules. Multiple monoclonal antibodies raised against epithelial antigens have helped identify a range of distinct, novel protein epitopes.


OBJECTIVE
In this study, we raised a monoclonal antibody to detect a novel epithelial molecular component.


METHODS
We have produced a mouse monoclonal antibody using normal human amniotic tissue as an immunogen. The monoclonal antibody was subsequently immunohistochemically screened, and the target antigen was cloned using an immunoscreening method.


RESULTS
In the course of the screening, we identified unique antibody staining patterns within the cytoplasm of a subset of amniotic cells at intervals within the normal placental epithelia. By immunoscreening, we identified this candidate gene as laminin receptor (LR). By dot blot analysis, this antibody reacted with recombinant LR. The same localization of the antigen and LR was proved by a double staining immunofluorescence test in the placenta. This monoclonal antibody unexpectedly demonstrated linear staining within the dermal-epidermal junction of normal human skin but failed to react within the keratinocyte cytoplasm.


CONCLUSIONS
We have produced and characterized a novel monoclonal antibody 29A that recognizes an LR-related molecule, which demonstrated a unique staining pattern. This monoclonal antibody might be a useful tool for further investigations into the epithelial tissues and the cutaneous basement membrane (BM).
Ota
Mitsuhito
M
Department of Dermatology, Hokkaido University Graduate School of Medicine, N-15, W-7, Sapporo 060-8638, Japan.
Sawamura
Daisuke
D
Yokota
Koichi
K
Ueda
Masamichi
M
Horiguchi
Yuji
Y
Kodama
Kazuo
K
Goto
Maki
M
Shimizu
Hiroshi
H
D000650:Amnion / Q000737:chemistry*; D000911:Antibodies, Monoclonal; D001485:Basement Membrane / Q000737:chemistry; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000737:chemistry; D004847:Epithelial Cells / Q000737:chemistry; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D011247:Pregnancy; D017474:Receptors, Laminin / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D012867:Skin / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D017474:Receptors, Laminin