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HTRF Human CCL5 Detection Kit, 500 Assay Points
HTRF Human CCL5 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
| Feature | Specification |
|---|---|
| Application | Protein Quantification |
| Sample Volume | 16 µL |
Product variants
Unit Size: 500 assay points
Part #:
62HCCL5PEG
Unit Size: 10,000 assay points
Part #:
62HCCL5PEH
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, and disposal requirements under European REACH regulations (EC 1907/2006).
HTRF Human CCL5 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
HTRF Human CCL5 Detection Kit, 500 Assay Points
Product information
Overview
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.
How it works
Assay principle
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.
Assay data analysis
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.
Assay details
Technical specifications of human CCL5 kit
| 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 |
Analytical performance
Intra and Inter assay
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% |
Assay validation
CCL5 secretion in PBMCs stimulated with LPS, IL1ß and TNFa
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.
CCL5 secretion in LX2 stimulated with TGFß
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.
Specifications
| Application |
Protein Quantification
|
|---|---|
| Brand |
HTRF
|
| Detection Modality |
HTRF
|
| Product Group |
Kit
|
| Sample Volume |
16 µL
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Target Class |
Cytokines
|
| Target Species |
Human
|
| Technology |
TR-FRET
|
| Therapeutic Area |
Metabolism/Diabetes
NASH/Fibrosis
Oncology & Inflammation
|
| Unit Size |
500 assay points
|
Video gallery
HTRF Human CCL5 Detection Kit, 500 Assay Points
HTRF Human CCL5 Detection Kit, 500 Assay Points
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