General Block ELISA Blocking Buffer

If fluorescence-stained cells cannot be evaluated immediately after labeling with the fluorescence probes, add Fixative to the cells at a ratio of 1:10. For example, add 100 µL Fixative to 900 µL cells and incubate for at least 15 minutes at RT in the dark. Never add Fixative until the staining and final wash steps have been completed. Fixed cells may be stored on ice or at 4°C up to 24 hours.

Reduces background using a mixture of blocking agents, including BSA. General Block contains mammalian protein blocking agents to provide adequate blocking strength for most immunoassays, including monoclonal and polyclonal antibody capture ELISAs and peptide and protein antigen-down ELISAs. This unique blocking buffer contains a heterogeneous mixture of proprietary protein stabilizers and small molecules (including BSA) that block the uncoated regions of the plate. Blocking with ICT’s General Block minimizes non-specific binding interactions during the assay process to reduce background noise and enhance the sensitivity of the assay. General Block provides a microhydrated environment to stabilize the adsorbed protein. This prevents degradation of the coated material and improves retention of protein antigenicity or antibody activity during long-term storage. General Block contains an antimicrobial agent for room temperature blocking of the plate and for long-term storage of the dried plate at 2-8°C. When preparing plates, the antibody or antigen is typically coated using 50-200 µL of coating solution per Well. After coating, plates are normally washed to remove unbound proteins and then blocked using a larger volume of blocking buffer than was used for coating, such as 300 µL per Well. This ensures that all uncoated regions inside the Well are blocked. A 96-Well plate blocked using this method will require 28.8 mL of blocking solution. However, allow approximately 10% extra blocking buffer to account for losses during pipetting.