Human CCL3 (MIP1 alpha) kit HTRF®

Name: Human CCL3 (MIP1 alpha) kit
Brand: Cisbio
SKU: 40394
Availability: InStock

Associated products

The HTRF human CCL3 (MIP1 alpha) kit is designed for the quantification of human CCL3 release in cell supernatant.

No-wash
Under 2h bench time
Highly specific

Download the protocols

The HTRF human CCL3 (MIP1 alpha) kit is designed for the quantification of human CCL3 release in cell supernatant.

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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.

Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma. Check out the Humant CCL3/MIP-1α AlphaLISA kit’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples.

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.

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.

Can HTRF technology answer to different scientific questions? An academic with industrial standards point of view

In collaboration with CNRS - Scientific Presentations

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Monitor immune regulation the simple way - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the Immune System - Brochures

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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

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

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Portfolio of Life Sciences Cytokine Assays

This leaflet presents the analytical performances of each assay - Brochures

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Get the brochure about technology comparison. - Brochures

Safety Data Sheet (DEU) CCL3 (h) Kit / 62HCCL3PEG