When developing any new ELISA, it is often helpful to screen several different sample diluent formulations to better match the matrix complexity present in the sample wells with the standard or calibrator wells. Most of the time invested in developing a new ELISA will involve selecting the proper calibrator matrix that matches most closely to the often very complex biological sample matrix. Comparing four different sample diluent formulations on the same IgG or antigen-coated ELISA plate makes it easy to determine which diluent will best equalize antibody-binding kinetics between the sample and calibrator wells. The Sample Diluent Optimization Pack provides 100 mL of each of our four Sample Diluent formulations to begin optimizing the sample diluent portion of the assay development process.
Sample Diluent Optimization Pack
ICT’s Sample Diluent Optimization Pack provides four 100 mL bottles of Sample Diluent formulations (General Serum Diluent, Plasma Sample Diluent, Neptune Sample Diluent, and Protein-Free Sample Diluent) for an economical and fast method to screen for the best Sample Diluent for an assay.
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To quickly determine which Sample Diluent formulation is best for the assay, all four Sample Diluents can be run in parallel on the same IgG-coated ELISA plate.
1. Coat antibody or antigen onto the ELISA plate using ICT’s Antibody Coating Buffer or Antigen Coating Buffer.
2. Incubate 8–24 hours at room temperature (RT).
3. Aspirate the coating solution.
4. Wash each well twice with ICT’s ELISA Wash Buffer.
5. Block the uncoated regions of the ELISA plate by pipetting 300–400 µL of block buffer into all wells and incubate 8–24 hours at RT.
6. Designate wells for the evaluation of each Sample Diluent formulation, and then prepare standards and samples spiked with known amounts of target analyte in the Sample Diluents.
7. Load 100 µL of each sample or standard into appropriate wells.
8. Proceed with the rest of the ELISA protocol.
9. Analyze results to determine which Sample Diluent formulation gave the lowest background signal, highest signal-to-noise ratio, and most accurate recovery of any samples spiked with known amounts of analyte.
• pH: 7.4 at 1X
• Contains: General Serum Diluent – Mammalian proteins
Plasma Sample Diluent – Mammalian proteins
Neptune Sample Diluent – Non-mammalian proteins
Protein-Free Sample Diluent – Synthetic molecules
• Supplied At: 1X
• Storage: 2-8°C