Immunoelectron microscopy shows that all of the LH7.2 binding sites are localized at the inferior border of the lamina densa. No binding could be detected within the dermis underneath the basement membrane at the putative sites of anchoring plaques. The antibody reacts predominantly with the basement membrane zone of stratified squamous epithelia and ectodermally derived glands. No staining is present around "simple" glandular epithelia or around blood vessels, muscle and nerve fibers. Detailed staining patterns of antibody LH7.2 with Type VII collagen in various normal and malignant tissues have been described (1). There is no loss of staining by treatment of frozen tissue sections with enzymes such as bacterial collagenase, trypsin, pronase, neuraminidase, chondroitinase or hyaluronidase. Staining is lost by pepsin digestion and by fixation with methanol, and is reduced by formaldehyde and glutaraldehyde fixation.

Suitable for use in Immunofluorescence, ELISA, Dot Blot, Western Blot, Immunohistochemistry, Immunocytochemistry and Immunoelectron Microscopy. Other applications not tested.

Immunofluorescence (Indirect): 1:1000 on human or other mammalian frozen sections Immunohistochemistry (Frozen): 1:2000 on human tongue section fixed in acetone. Specifically stains basement membranes. Optimal dilutions to be determined by the researcher.

Secreted, extracellular space, extracellular matrix, basement membrane

Fusion of mouse myeloma cells and splenocytes from BALB/c mice

May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Insoluble fractions prepared from neonatal human foreskin epidermal cells.

Supplied as a liquid, 15mM sodium azide.

Ascites

Recognizes human tissue Type VII collagen. Recognizes an epitope located on collagenase digested Type VII collagen (150kD), i.e., the non-helical carboxy terminal region of the Type VII collagen dimer. It is also reactive in immunoblotting against intact tissue Type VII collagen (250kD), but once the terminal regions are completely digested by pepsin, this reactivity is completely lost. No reactivity against collagen types I, II, III, IV, V, pepsin-digested collagen type VII, laminin and fibronectin. Species Crossreactivity: Basement membrane zone of stratified squamous epithelia of human, bovine, porcine, goat, guinea pig, monkey, marmoset and sheep.