DiaPharma Group, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

Reagents -- Composition, Packaging and Storage

1. Capture Antibody (Catalog No. RBFG-EIA-C): Yellow capped tube containing 0.4 ml capture IgG affinity-purified sheep anti- rabbit Fibrinogen (Fg) IgG for coating ELISA plates.
2. Detecting Antibody (Catalog No. RBFG-EIA-D): Red capped tube containing 0.4 ml IgG HRP-conjugated affinity purified sheep anti-rabbit Fibrinogen (Fg) for detection.

Both reagents should be stored at -20°C.

Procedure:

1. Coating of plates: Dilute the capture antibody 1/100 in coating buffer and immediately add 100 ul to each well of the microplate. Seal the plate or place it in a humid chamber to prevent drying. Incubate 2 hrs @ 22°C or overnight @ 4°C. If not contaminated the sealed plates can be kept at 4°C for one week.
2. Blocking: Blocking is not required under the conditions described. Washing the plate four times with PBS-Tween is sufficient to block non-specific interactions.
3. Samples: Dilutions of standard plasma are made in a range from 1/10,000 (100%) down to 1/320,000 (3.25%). Test plasma samples are diluted 1/20,000 & 1/40,000. All dilutions are made in PBS-Tween. Apply 100 microliters/well and incubate plate @ 22°C for 60 minutes. Wash X 4 with PBS-Tween.
4. Detecting Antibody: Dilute the detecting antibody 1/100 in HBS-Tween-BSA diluent and apply 100 microliters to each well. Incubate plate @ 22°C for 60 minutes. Wash X 4 with PBS-Tween.
5. 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:

• SARFG-IG - Sheep anti-rabbit Fibrinogen, purified whole IgG from antiserum 10 mg vial
• SARFG-AP - Sheep anti-rabbit Fibrinogen, affinity purified IgG 0.5 mg vial
• SARFG-HRP - Sheep anti-rabbit Fibrinogen, peroxidase labelled whole IgG 0.2 mg vial