Mouse CXCL12/SDF-1 alpha Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse SDF-1 alpha Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse SDF-1 alpha in cell culture supernates and mouse serum. It contains E. coli-expressed recombinant mouse SDF-1 alpha and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant mouse SDF-1 alpha. Results obtained using natural SDF-1 alpha 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 of natural mouse SDF-1 alpha.

Preparation and Storage

Background: CXCL12/SDF-1 alpha

CXCL12/Stromal Cell-derived Factor 1 (SDF-1) is a member of the CXC chemokine ligand superfamily. Six alternatively spliced CXCL12/SDF-1 isoforms have been identified including, CXCL12/SDF-1 alpha, CXCL12/SDF-1 beta, and CXCL12/SDF-1 gamma. CXCL12/SDF-1 alpha is 89 amino acids (aa) in length with a predicted molecular weight of 10 kDa, CXCL12/SDF-1 beta is 93 aa in length with a predicted molecular weight of 10.6 kDa, and CXCL12/SDF-1 gamma is 119 aa in length with a predicted molecular weight of 13.7 kDa. The mouse CXCL12/SDF-1 orthologs share 99% aa sequence identity with the human protein. The alpha and beta isoforms are ubiquitously expressed while the gamma isoform is primarily expressed in the heart. CXCL12/SDF-1 isoforms are chemotactic for T lymphocytes and monocytes. It is cardio protective and has also been shown to hinder CXCR4-mediated HIV infection of T cells.

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.