Human Carbonic Anhydrase IX Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Carbonic Anhydrase IX Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CA9 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human CA9 ectodomain and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CA9 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human CA9.

Preparation and Storage

Background: Carbonic Anhydrase IX/CA9

Carbonic anhydrases catalyze the reversible reaction of CO2 + H2O = HCO3- + H+, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption. Human members of the CAs are secreted (CA6), membrane-bound (CA4, 9 to 12 and 14), cytosolic (CA1 to 3, 7, 8 and 13) or mitochondrial (CA5A and 5B). Some CAs may serve as markers for tumors and hypoxia. A subset of CAs lack CA activity due to point mutations but retain esterase function.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1、Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2、Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3、Add 50 μL of Assay Diluent to each well.

4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.