INTENDED USE
The Vet Insulin RIA Test Kit has been developed for the determination of insulin in animals. The kit has been validated on and normal ranges have been established for canine, feline, and equine species.
ANALYTICAL PRINCIPLE
Insulin from samples and standards along with insulin from 125I-Insulin Tracer are allowed to compete for binding to the insulin antibody for 20 hours. After incubation, 2nd antibody is added to the tubes to capture the (125I)insulin-antibody complex. The (125I)insulin-antibody-2ndantibody complex is precipitated in the tubes by the addition of a precipitating solution. The tubes are then centrifuged to allow the collection of the complexes at the bottom of the tube. The liquid layer that contains unbound 125I-Insulin Tracer is decanted from the tubes. The tubes are read on a gamma counter following the decanting step. The level of insulin in the samples and standards are inversely related to the level of radioactivity that is measured.
PROCEDURE
- Label the required number of glass tubes according to the Tube Assignment Table below.
- Dispense 200 µL of B0 (C) to tubes 3 – 4.
- Dispense 100 µL of B0 (C), Standards (C1 – C6), and samples to tubes 5 – end.
- Dispense 100 µL of 1st Antibody (A) to tubes 5 – end.
- Dispense 100 µL of 125I-Insulin Tracer (B) to all tube.
- Vortex tubes 3 – end.
- Cover the tubes with Parafilm.
- Incubate the tubes for 20 hours + 5 minutes at room temperature (15 °C – 25°C).
- Dispense 50 µL of 2nd Antibody (D) to tubes 3 – end.
- Vortex tubes 3 – end.
- Incubate the tubes for 15 minutes + 30 seconds at room temperature (15 °C – 25°C).
- Dispense 1 mL of Precipitation Solution (E) to tubes 3 – end.
- Vortex tubes 3 – end.
- Incubate the tubes for 15 minutes + 30 seconds at room temperature (15 °C – 25°C).
- Centrifuge tubes 3 – end at 3,500 RPM for 30 minutes + 1 minute.
- Decant or aspirate all tubes.
- Count the tubes with a gamma counter for 1 minute.
Inquire at info@ivdtechnologies.com for the normal range, glucose curve, and validation study.