For Research Use Only. Not For Use In Diagnostic Procedures.
Reagents -- Composition, Packaging and Storage
- Capture Antibody (FII-EIA-C): One yellow-capped vial containing 0.5 ml of polyclonal affinity purified anti-FII antibody for coating plates.
- Detecting Antibody (FII-EIA-D): One red-capped vial containing 0.5 ml of peroxidase conjugated polyclonal anti-FII antibody for detection of captured FII.
Note: Antibodies are supplied in a 50% (v/v) glycerol solution for storage at -10 to -20°C. Keep vials tightly capped. Do not store in frost-free freezers.
Assay Procedure:
- Coating of plates: Dilute the capture antibody 1/100 in coating buffer (preferably in a polypropylene tube) and immediately add 100 microliters to every well in the plate. Incubate 2 hrs @ 22°C.
- Blocking: Blocking is not required under the conditions described. Washing the plate with PBS-Tween is sufficient to block nonspecific interactions. Wash plate X 3 with wash buffer.
- Samples: Reference plasma is diluted 1/5,000 (100%) then serial 1/2's down to 1/160,000 (3.13%). Sample plasmas are diluted 1/10,000, 1/20,000 & 1/40,000. All dilutions are made in HBS-BSA-T20 sample diluent. Apply 100 microliters/well and incubate plate @ 22°C for 60 minutes. Wash plate X 3 with wash buffer.
- Detecting Antibody: Dilute the detecting antibody 1/100 in HBS-BSA-T20 sample diluent and apply 100 microliters to each well. Incubate plate @ 22°C for 60 minutes. Wash plate X 3 with wash buffer.
- OPD Substrate: Apply 100 microliters of freshly prepared OPD substrate to every well. Allow color to develop for 10-15 minutes then stop color reaction with the addition of 50 microliters/well of 2.5 M H2SO4. The plate can be read at a wavelength of 490 nm.
Related Products:
- SAFII-IG - Sheep anti-human FII, IgG from antiserum
- SAFII-AP - Sheep anti-human FII, affinity-purified IgG
- SAFII-HRP - Sheep anti-human FII, IgG-peroxidase conjugate
- FII-EIA - Paired antibody set for ELISA of FII, 5 x 96 well
- FII-DP - Human plasma deficient in FII, immune depleted