Human CD117/c-kit Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human SCF sR Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human SCF sR in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human SCF sR and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human SCF sR 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 SCF sR.

Preparation and Storage

Background: CD117/c-kit

CD117, also known as c-kit and SCF R, is the cellular receptor for Stem Cell Factor (SCF), also named c-kit ligand, mast cell growth factor, and steel factor. This receptor-ligand system plays essential roles in germ cell development, melanogenesis, hematopoiesis, and oncogenesis. CD117 is expressed on lymphoid, myeloid, erythroid, and megakaryocyte progenitors, NK cells, germ cells, melanocytes, glial cells, vascular smooth muscle cells, placenta, and epithelial cells. Altered levels and mutations in CD117 are associated with several types of cancer including lung, breast, gastrointestinal stromal, and germ cell tumors.

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.