The Human CCL3 (MIP1 alpha) kit is designed for the quantification of human CCL3 release in cell supernatant.
See more
  • No-wash No-wash
  • Under 2h bench time Under 2h bench time
  • Highly specific Highly specific
The Human CCL3 (MIP1 alpha) kit is designed for the quantification of human CCL3 release in cell supernatant.
-
In stock

Overview

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

Benefits

  • NO CROSS WITH CCL4 OR 5
  • VALIDATED ON PBMC

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 Cisbio 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.
CCL3 kit assay principle

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.

To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting, please visit this page.

Cisbio also worked with Myassays.com to help you in your data analysis. By clicking on this link, you’ll be able to access a free online software to run your CCL3 analysis.

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

Intra and Inter assay

intra assay (n=24)

Sample Mean [CCL3] (pg/ml) cv
1 133 9%
2 667 4%
3 3,450 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 2,316 4682 4%
Mean CV 5%

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 Cisbio 96-well low volume white plate to be analyzed by the Human CCL3 kit.
CCL3 secretion in PBMC stimulated with LPS, IL1ß and TNFa
HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the immune system - Brochures

Normalize HTRF cytokine assays with viability assay on Spectramax I3X-Minimax

In collaboration with Molecular Devices - Application Notes

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Application Notes

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Technical Notes

How to freeze PBMC

Guideline for PBMC freezing - Technical Notes

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

Product Insert CCL3 (h) Kit / 62HCCL3PEG-62HCCL3PEH

62HCCL3PEG-62HCCL3PEH - Product Insert

HTRF makes cytokine detection easy

A fun video introducing you to cytokines assays with HTRF - Videos

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

tab6