General Serum Diluent

1. Dilute ELISA Wash Buffer, 10X by adding 1 part ELISA Wash Buffer to 9 parts deionized water and mix for 15 minutes. ELISA Wash Buffer, 10X is supplied concentrated, so crystalline precipitates may form in the bottle, especially when refrigerated. If this happens, gently warm or mix the buffer until all crystals are dissolved. Do not let it boil., 2. Completely fill the Wells of the ELISA plate with 1X wash buffer (about 400 µL/Well). The diluted wash buffer may be dispensed through a squirt bottle, a plate washer, through a multi-channel pipette, or through an automated system., 3. Aspirate or dump out the buffer and repeat for a total of 2-4 washes., 4. After the final dump or aspiration, pound the plate on paper towels to remove any excess liquid., If washing the plate during the coating process, we recommend 2-3 washes after the coating step before adding the block buffer. If washing the plate as part of an ELISA procedure, we recommend 3-4 washes after the sample incubation and after the conjugate incubation.

A stable protein matrix for dilution of serum and other aqueous-based biological samples. General Serum Diluent is formulated to provide a stable, protein-friendly environment for dilution of serum, cell culture, ascites, urine, and other aqueous-based biological samples for evaluation within an ELISA format. It can serve as an excellent generic matrix in which to dilute antigen standards when preparing standard curves within antibody-sandwich formats. It also provides an excellent serum dilution medium for antigen-down ELISAs in the assessment of a humoral immune response to a particular disease state or immunization regimen. Sample diluents are used to dilute samples into the functional range of the assay and to create the standard curve. Due to the finite binding capacity of the plate Well-coated proteins (e.g., antibodies, antigens), highly concentrated samples must be diluted in order to obtain absorbance readings within the sensitivity detection limits of the instrument. Properly formulated sample diluents will also reduce background noise associated with non-specific bridging of signal-generating conjugates to the plate Well surface. General Serum Diluent provides a BSA protein-buffered, neutral pH environment that is highly compatible with antibody-antigen interactions. Inclusion of a protein component helps minimize the degree of non-specific IgG adsorption onto coated and blocked ELISA plate Wells during antigen-down screening of serum or plasma samples. Antimicrobial agents allow for room temperature bench-top use and extensive storage stability at 2-8°C.