Human Attractin Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Attractin Immunoassay is a 4.5 hour solid phase ELISA designed to measure Attractin levels in cell culture supernates, serum, plasma, saliva, and urine. It contains CHO cell-expressed recombinant human Attractin and antibodies raised against the recombinant protein. Results obtained for naturally occurring human Attractin showed linear curves that were parallel to the standard curves obtained using the Quantikine Human Attractin Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human Attractin.

Preparation and Storage

Background: Attractin

Attractin/ATRN, also known as DPPT-L, is an approximately 200 kDa transmembrane glycoprotein that shows dipeptidyl peptidase activity similar to DPPIV/CD26. Attractin is involved in a variety of processes including monocyte-T cell adhesion, axon myelination, melanocyte pigment synthesis, and energy homeostasis. Attractin is transiently upregulated during T cell activation before expression switches to the 175 kDa secreted isoform which is released into the circulation. Soluble Attractin is preferentially expressed by leukocytes and differentiating neurons. It blocks neurite formation and is elevated in the CSF of astrocytoma patients.

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.