Human IFNAR2(Interferon alpha/beta receptor 2) ELISA Kit

Matrices listed below were spiked with certain level of IFNAR2 and the recovery rates were calculated by comparing the measured value to the expected amount of IFNAR2 in samples.

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of IFNAR2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Human IFNAR2(Interferon alpha/beta receptor 2) ELISA Kit components

96 Assays

Storage

Serum

Use a serum separator tube (SST) and allow samples to clot for 30 mins before centrifugation for 15 mins at ~1000 × g. Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C.

Plasma

Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles. Note: over hemolyzed samples are not suitable for use with this kit.

Urine & Cerebrospinal Fluid

Collect in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and if any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.

Cell culture supernatant

Collect supernatant and centrifuge at 4°C for 20 mins at 2000-3000 rpm. Remove supernatant and rinse cells twice with PBS (pH 7.2-7.4) and perform a total cell count. Optimal cell concentration is 1 million/ml. To release cellular components, dilute the cell pellet in PBS and use 3-4 freeze-thaw cycles in liquid Nitrogen (commercial lyses buffers can be used according to manufacturer’s instructions). Centrifuge at 4°C for 20 mins at 2000-3000 rpm to pellet debris and remove clear supernatant to clean microcentrifuge tube for ELISA or storage.

Tissue Homogenates

As hemolysis blood can have an impact on assay result, it is necessary to remove residual blood by washing the tissue with pre-cooling PBS buffer (0.01M, pH=7.4).Mince tissue after weighing it and homogenize in PBS (the volume depends on the weight of the tissue. Usually, 9mL PBS would be appropriate for 1 gram tissue pieces. Some protease inhibitors are recommended to add to the PBS) with a glass homogenizer on ice. To further breakdown the cells, you can sonicate the suspension with an ultrasonic cell disrupter or subject it to freeze-thaw cycles. The homogenates are then centrifuged for 5 minutes at 5000×g in order to get the supernatant. The total protein concentration was determined by BCA kit and the total protein concentration of each pore sample should not exceed 0.3mg.