carbonate buffer elisa
elisa coating buffer ph
elisa plate coating buffer
elisa coating buffer sodium bicarbonate
pbs as coating buffer for elisa
elisa coating buffer pbs
elisa carbonate coating buffer
carbonate coating buffer
antibody coating buffer
elisa plate coating
coating elisa plates at 37 degrees
antibody coating on elisa plate
elisa coating plate
coating in elisa
coating antibody for elisa
elisa antibody coating
elisa peptide coating
To immobilize the antigen or antibody on the solid surface (of ELISA or FLISA plate) the pH of the coating buffer is critical for proper immobilization of the "catcher" protein or peptide at the charged surface of the plate. The pH may significantly affect both the level of molecule adsorption, as well as its structure (this is equally true for both peptides and proteins, especially antibodies).
Therefore, we offer two coating buffers, Immobilizer pH7.4 and pH9.6. One of them will be perfect for your assay.
For optimal coating of the pate we strongly recommend testing both buffers side by side at the first stage of the assay development. Depending on the brand of plates you are using, as well as the nature of the protein or peptide, the optimal time of incubation of the "catcher" solution in your plate may very significantly. Thus we recommend starting with the overnight incubation of molecules in plate at +4 to 8 degrees, and, consequently, optimize the coating procedure for every type of absorbed molecule individually.
Please note that the Immobilizer buffers are supplied as 10x concentrate, so before use simply dilute the stock with deionized water.
Immobilizer 7.4 10x 125 ml #AP0510-125
500 ml #AP0510-500
Immobilizer 9.6 10x 125 ml #AP0511-125
500 ml #AP0511-500
Using coating Buffers (general guide)
- Dilute protein, peptide or antibody you want to absorb in Immobilizer buffer in concentration 0.1 - 10 μg/ml
- add 100-150 μl/well if you use 96-well plate, for 384-well plate adjust the volume accordingly. In general we recommend filling the well for 50% of total well volume
- Incubate the plate (closed or sealed) for 1-4 hours at room temperature or at 4°C overnight. The buffer does not contain any components that could affect immobilization of your preparation
- Wash the pate 3-5 times with 300 μL of Plate Wash Buffer ((1x concentration) The buffer is supplied as 10x stock, do not forget to dilute it to the working concentration)).