Figure 2 Immunofluorescence methods. (A) Direct immunofluorescence (DIF) method uses fluorescein-labeled antibodies (IgA, IgG, IgM) to immunoglobulins or complement that is deposited in the BMZ area in patient's skin. (B) Indirect immunofluorescence (IIF) method uses fluorescein-labeled antibodies to immunoglobulins to test if patient's blood contains circulating antibodies that will bind to BMZ molecules in normal skin. (C) Salt split skin (SSS) method uses normal or patient's skin treated with 1 M NaCl to separate epidermis and dermis in the lamina lucida. Labeled antibodies determine if antigens are on the epidermal or dermal side of the split. In this example, bullous pemphigoid antigen (BP) is part of the hemidesmosome, and is found on the epidermal side of the artificial blister induced by 1 M NaCl.
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