Uniprot No.: P01308
Species: Homo sapiens (Human)
Sample Types: serum, plasma, cell culture supernates, tissue homogenates
Detection Range: 10 μIU/ml-160 μIU/ml
Sensitivity: 2 μIU/ml
Assay Time: 1-5h
Sample Volume: 50-100ul
Detection Wavelength: 450 nm
Assay Principle: quantitative
Measurement: Sandwich
Materials provided:
- An assay plate --- The 96-well plate is composed of 12 x 8 strip plates and can be dismountable.
- Six bottles of lyophilized standard --- Reconstitute one bottle of the standard with sample diluent at dilution series and draw the standard curve.
- Biotin-antibody (100 x concentrate) 1 x 120 μl --- Act as the detection antibody and need to be diluted before use.
- HRP-avidin (100 x concentrate) 1 x 6 ml--- Catalyze the TMB substrate for color development and need to be diluted before use.
- Biotin-antibody Diluent 1 x 15 ml --- Dilute the Biotin-antibody.
- HRP-avidin Diluent 1 x 15 ml --- Dilute the HRP-avidin.
- Sample Diluent 1 x 50 ml --- Reconstitute the standard and dilute the sample to an appropriate concentration.
- Wash Buffer (20 x concentrate) 1 x 15 ml --- Wash away the unbound solution and non-specific substances.
- Substrate A 1 x 7 ml --- Contain oxidant.
- Substrate B 1 x 7 ml--- Contain TMB. TMB interacts with HRP, eliciting the solution turns blue.
- Stop Solution 1 x 10 ml --- Add stop solution and stop the color development. The solution color immediately turns from blue to yellow.
- Adhesive Strip (For 96 wells) 4 --- Cover the microtiter plate when incubation.
- An instruction manual
Materials not provided:
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 600 nm or 630 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution