Material not included
- Coating Buffer: PBS (Phosphate Buffered Saline), pH 7.4, 12 mL is sufficient for 96 tests.
- Wash Buffer: PBS with 0.05 % (v/v) Tween-20, 500 mL is sufficient for 96 tests.
- Blocking Buffer: Wash Buffer with 2 % (w/v) bovine serum albumin, 35 mL is sufficient for 96 tests.
- Dilution Buffer: Wash Buffer with 0.5 % (w/v) bovine serum albumin, 50 mL is sufficient for 96 tests.
- Substrate Stock: Solution 10 mg/mL TMB in Dimethyl sulfoxide, 1 mL is sufficient for 96 tests. Protect from light.
- Substrate Dilution Buffer: 50 mM disodium hydrogen phosphate (Na2HPO4) and 25 mM citric acid, adjust pH to 5.5 with 1 M Sodium hydroxide (NaOH), 25 mL is sufficient for 96 tests.
- TMB Substrate Working Solution:
For each plate dilute 250 μL substrate stock solution in 25 mL substrate dilution buffer and add 12 μL 5 % H2O2 (pipette 10 μL 30 % H2O2 into 50 μL distilled water), mix well.
1) The TMB Substrate Working Solution should be freshly prepared and used within 15 minutes.
2) If you choose to use other commercially available ready-to-use TMB substrate solutions, you should follow the manufacturer's instruction
- Stop Solution: 1 M sulfuric acid (aqueous), 6 mL is sufficient for 96 tests.
- High binding surface 96-well microplate, clear flat bottom
- Microplate sealing film
- Pipettes and pipette tips
- UV/Vis microplate spectrophotometer (absorbance 450nm, correction wavelength set to 600 nm).