| | Re: emeargancy helppppp
Protein-Free Blocking Buffer is used for reduces or eliminates many of the problems encountered with traditional protein-blocking reagents, such as cross-reactivity and interference from glycosylation. When to use protein-Free Blocking Buffers with Tween-20 it has to be high purified free of peroxides and carbonyls that may interfere in some immunoassay systems. The Protein-Free Blocking Buffers may be used as a protein stabilizer for drying antigen- or antibody-coated microplates and, before completely dry plate, sealing in a plastic bag with desiccant. Store plate at 4°C.
TBS (10-50 mM Tris-HCl buffer) 0.15 M NaCl at pH 7.4 with or without 0.05 % Tween-20, 1-5 % sucrose, 1 % glucose, 0.02-0.15 % Kathon CG. Store unopened product at room temperature. After opening, store product at 4°C.
PBS (0.15 M PO4 buffer) at pH 7.4 with or without 0.05 % Tween-20, 1-5 % sucrose, 1 % glucose, 0.02-0.15 % Kathon CG. Store unopened product at room temperature. After opening, store product at 4°C.
Procedure for blocking ELISA plates : coat the ELISA plate with antigen or antibody according to standard procedures. Add 300 μl of the Protein-Free Blocking Buffer, better that without Tween-20, to each well and incubate for 1 hour at room temperature. Alternatively, add 300 μl of blocking buffer to each well and immediately invert plate to empty contents. Repeat this process two more times. From that moment proceed with assay or invert plate, and allow it to completely dry for about 2 hrs. Place dry plate in a plastic bag or other container with desiccant and store at 4°C.
Procedure for blocking membranes : add sufficient Protein-Free Blocking Buffer containing 0.05 % Tween-20, to cover the entire surface of the membrane. Incubate for 1 hour at room temperature on a rocking platform at low speed. Continue the blotting procedure using the Protein-Free Blocking Buffer to dilute primary and secondary antibodies.