The HTRF human CCL5 (RANTES) kit is designed for the quantification of human CCL5 release in cell supernatant.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
CCL5, also known as RANTES (regulated upon activation, normal T-cell expressed and secreted), is a chemokine mainly secreted by T lymphocytes, macrophages, platelets, and certain types of tumor cells. RANTES plays an important role in inflammatory sites by recruiting T cells and macrophages. When secreted with IL-2 and IFN-γ, RANTES also induces activation and proliferation of natural killer (NK) cells. function of RANTES in oncology is still unclear, but in infectious diseases RANTES has been described as inhibiting HIV entry into target cells.
Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable detection of low levels of analytes in serum or plasma.
Assay Points |
500
|
---|---|
Assay Target Type |
Kit
|
Assay Technology |
HTRF
|
Brand |
HTRF
|
Quantity |
1
|
Shipping Conditions |
Shipped in Dry Ice
|
Therapeutic Area |
Metabolism/Diabetes
NASH/Fibrosis
Oncology & Inflammation
|
Unit Size |
500 Assay Points
|
Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Revvity low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.
The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.
Revvity also worked with Myassays.com to help you in your data analysis.
Sample size | 16 µL |
---|---|
Final assay volume | 20 µL |
Kit components | Lyophilized standard, frozen detection antibodies, buffers &protocol |
LOD &LOQ (in Diluent) | 7 pg/mL &18 pg/mL |
Range | 18 2,000 pg/mL |
Time to result | ON @ RT |
Calibration | NIBSC (92/520) value (IU/mL) = 0,015 x HTRF hCCL5 value (pg/mL) |
Species | Human only |
Intra assay (n=24)
Sample | Mean [CCL5] (pg/mL) | CV |
---|---|---|
1 | 74 | 7% |
2 | 417 | 7% |
3 | 1044 | 6% |
Mean CV | 7% |
Inter assay (n=4)
Sample | [CCL5] (pg/mL) | Mean (delta ratio) | CV |
---|---|---|---|
1 | 62 | 320 | 12% |
2 | 256 | 1214 | 13% |
3 | 1008 | 4405 | 5% |
Mean CV | 10% |
PBMCs were plated at 100,000 cells/well in a 96 well plate, and stimulated for 24h with LPS, IL1ß and TNFa respectively used at 200 µg/mL, 10 ng/mL and 10 ng/mL. Supernatants were diluted to 1/3, and 16 µL of the diluted samples were transferred into a 384 well (SV) white detection plate to be analyzed by with the Human CCL5 Assay.
LX2 cells were plated at 400,000 cells/well in a 6 well plate, cultured for 3 days, then starved for 1 day before 48 hours of stimulation with TGFß at 0.5ng/ml. Next, 16 µL of supernatant were transferred into a 384 well (SV) white detection plate to be analyzed with the Human CCL5 Assay.
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