human recombinant type vii collagen  (Lotus Tissue Repair Inc)

Journal:

Article Title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells

doi: 10.1111/j.1365-2249.2006.03115.x

Figure Lengend Snippet: Generation of recombinant mouse type VII collagen NC1 domain and polyclonal antibodies against mouse NC1. (a). The cDNA of mouse NC1 domain encoding 580 amino acids at the N-terminal was cloned and expressed in Escherichia coli with a molecular mass of approximately 86 kD (lane 2, Coomassie blue-stained). The recombinant protein has a V5 epitope which is used for identifying the correct recombinant protein production by antibody to V5 (lane 3). The serum from the purified NC1 recombinant protein-immunized rat reacts specifically with NC1 (lane 4), while pre-immune serum (same dilution) of the same rat does not react with the protein (lane 5); 2·4 µg, 1·2 µg, 0·12 µg and 0·12 µg recombinant NC1 were loaded to lanes 2, 3, 4 and 5, respectively. (b). The serum from the purified NC1 recombinant protein-immunized rat also reacts distinctively with skin basement membrane zone of the dermal–epidermal junction and hair follicles (right panel), but the pre-immune serum of the same rat does not label it (left panel).

Article Snippet: Cloning and generation of a recombinant mouse type VII collagen NC1 domain The first-strand cDNA synthesis was accomplished using mouse total skin RNA (Origene, Rockville, MD, USA) with a reverse transcription kit and Random Decamers primer (RETROscript kit) purchased from Ambion (Austin, TX, USA), according to the manufacturer’s instructions.

Techniques: Recombinant, Clone Assay, Staining, Purification