Description
CXCL12 (SDF-1) is a homeostatic chemokine constitutively expressed by bone marrow stromal cells and is present in many other tissues (skin, thymus, lymph nodes, lung, liver). It plays an essential role in homeostatic regulation of leukocyte traffic, hematopoiesis, organogenesis, cell differentiation and tissue regeneration. CXCL12 binds to and activates the CXCR4 receptor. CXCL12 and CXCR4 are overexpressed in various cancer types, and this aberrant expression strongly promotes proliferation, migration and invasion through multiple signal pathways.
Formats
Our 100 assay point kit allows you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
Features
No-wash steps, no separation steps
ELISA alternative technology
Broad sample compatibility
Small sample volume
Results in less than 2 hours
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Principle of the AlphaLISA CXCL12 assay
The AlphaLISA CXCL12 assay is based on an AlphaLISA sandwich immunoassay involving a biotinylated anti-CXCL12 antibody bound to Streptavidin-coated AlphaLISA Donor beads and an anti-CXCL12 antibody conjugated to AlphaLISA Acceptor beads. In the presence of the CXCL12 the beads come into proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer within the Acceptor beads, resulting in emission with λmax at 615 nm. The intensity of the signal is directly proportional to the concentration of CXCL12 present in the sample.
Protocol of the AlphaLISA human and mouse CXCL12 assay
The AlphaLISA CXCL12 assay can be run in a 96- or 384-well detection plate (50 µL final). As described here, samples or standards are dispensed directly into the assay plate for the detection of CXCL12 by AlphaLISA reagents. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally.
