ELISA

Overview

Enzyme-linked immunosorbent assay (ELISA) is a plate-based immunoassay used to quantify soluble proteins (antigens or antibodies) in biological samples such as conditioned medium, serum, or cell lysate. A capture antibody binds the target protein; a detection antibody conjugated to an enzyme produces a colorimetric or fluorescent signal proportional to analyte concentration.

Used by

• ELISA used to quantify soluble SEMA7A protein levels in conditioned medium from gallbladder fibroblasts cultured on soft (0.5 kPa) vs stiff (16 kPa) hydrogels, confirming that matrix stiffness drives secretion of SEMA7A as a paracrine factor PMID:24997986

Notes

• Sandwich ELISA (two-antibody format) provides higher sensitivity and specificity for complex biological matrices.
• ELISA of conditioned medium is a standard approach for quantifying secreted proteins in paracrine signaling studies.
• Results are typically expressed as ng/mL or pg/mL relative to a standard curve.

Sources

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