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HTRF Human CCL3 Detection Kit, 500 Assay Points

The HTRF human CCL3 (MIP1 alpha) kit is designed for the quantification of human CCL3 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 and disposal requirements under European REACH regulations (EC 1907/2006).

Feature Specification
Application Protein Quantification
Sample Volume 16 µL

The HTRF human CCL3 (MIP1 alpha) kit is designed for the quantification of human CCL3 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 and disposal requirements under European REACH regulations (EC 1907/2006).

Product Variants
Unit Size: 500 Assay Points
Part #:
62HCCL3PEG
List Price
USD 1,370.00
Unit Size: 10,000 Assay Points
Part #:
62HCCL3PEH
List Price
USD 14,590.00

Overview

Also called MIP-1 alpha (macrophage inflammatory protein 1 alpha), CCL3 is a member of MIP (macrophage inflammatory protein) family, which also includes CCL4 (MIP1 beta). This chemokine is secreted by monocytes, macrophages, T cells, and endolial cells. CCL3 plays an important role in activation of inflammatory response by attracting monocyte-lineae cells and lymphocytes to inflammation sites. CCL3 has been reported as a stem cell inhibitory factor.

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.

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
Oncology & Inflammation
Unit Size
500 Assay Points

Video gallery

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.

cytokines-how-it-works-assay-principle-human
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 CCL3 kit
Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol
LOD &LOQ (in diluent) 28 pg/mL &58 pg/mL
Range 58 4,400 pg/mL
Time to result ON @ RT
Calibration NIBSC (92/518) value (IU/mL) = 0.00005 x HTRF hCCL3 value (pg/mL)
Species Human only

 

Analytical performance

Intra and Inter assay

intra assay (n=24)

Sample Mean [CCL3] (pg/ml) CV
1 133 9%
2 667 4%
3 3450 4%
  Mean CV 6%

inter assay (n=4)

Sample [CCL3] (pg/ml) Mean (delta ratio) CV
1 178 298 8%
2 642 1138 3%
3 2316 4682 4%
    Mean CV 5%

 

Assay validation

CCL3 secretion in PBMC stimulated with LPS, IL1ß and TNFa

A dose response of LPS was performed on PBMC plated at 100,000 cells / well / 100 µL in a 96 well plate. Supernatants were diluted at 1/8th and 16µl of diluted samples were transferred into a Revvity 96-well low volume white plate to be analyzed by the Human CCL3 kit.

assay-validation-human-ccl3

 

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