After coating the ELISA plate with the capture antigen or antibody, proper blocking of the unoccupied areas of the plate wells is paramount to attaining an accurate signal. Assay sensitivity improves by utilizing a blocking buffer that effectively decreases background noise and improves the signal-to-noise ratio. Evaluation of block buffer candidates is an important step in the ELISA development process. The Block Buffer Optimization Pack provides five of ICT’s blocking buffer formulations for an economical approach to selecting the best blocking buffer formulation for an assay.
Block Buffer Optimization Pack
ICT’s Block Buffer Optimization Pack provides five 100 mL bottles of our different Blocking Buffer formulations (General Block, Neptune Block, SynBlock, Alternative Block, and Monster Block) for an economical approach to screening for the best blocking buffer for an ELISA.
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To quickly determine which Block Buffer formulation is best for the assay, all five Block Buffers can be screened in parallel on the same antibody or antigen-coated 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. Designate wells for the evaluation of each Block Buffer formulation, and then block the uncoated regions of the ELISA plate by pipetting 300–400 µL of each blocking buffer into the appropriate wells. Always use an equal or greater volume of blocking buffer than was used for the coating buffer solution.
6. Incubate 8–24 hours at RT. For best blocking, incubate overnight at RT.
7. Aspirate the blocking buffer; do not wash.
8. Proceed with the rest of the ELISA protocol.
9. Analyze results and select best block buffer based on which formulation gave the lowest background signal, highest signal to noise ratio, and most accurate recovery of any samples spiked with known amounts of antigen.
• pH: 7.4 at 1X
• Contains: General Block – Mammalian proteins
Neptune Block – Non-mammalian proteins
SynBlock – Synthetic molecules
Alternative Block – Synthetic molecules
Monster Block – Non-mammalian proteins
• Supplied At: 1X
• Storage: 2-8°C