Human CCL3 (MIP1 alpha) Kit

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

Cisbio's Human CCL3 kit is a homogeneous, add and read assay to measure CCL3 secretion in cell supernatant. It offers a fast and efficient alternative to ELISA.

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

Download Insider tips for PBMC isolation Technical Note
Download cytokine release from fresh blood samples application notes

Assay principle

The assay protocol, using a Cisbio 96-well low volume or a 384-well small volume white plate (20 µL final), is described below. Cell supernatant, sample or standard are dispensed directly into the assay plate for the detection by HTRF® reagents. 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 in 96 to 1536-well plates 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.

Add

Cytokines assay principle add

Read

Cytokines assay principle read

(1) Assay data analysis:

  • "Guidelines for data reduction" button will show you how to deal with HTRF data processing and 4PL 1/y2 fitting.
  • "MyAssays" button will direct you to a free online software able to run CCL3 analysis.

 

How to use HTRF cytokine assays

Analytical performance

Precision

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%

 

Assay performance

CCL3 secretion in PBMC stimulated with LPS, IL1β and TNFα

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 white HTRF HTRF 96-well low volume white plate to be analyzed by the Human CCL3 Assay

Human CCL3 assay kit secretion curve

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human CCL3 kit - 500 tests62HCCL3PEG
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Human CCL3 kit - 10,000 tests62HCCL3PEH
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Companion products

DescriptionCat. noProduct insertMSDS
Human CCL3 kit - 3 Standards62HCCL3CD3
-
Human CCL3 kit - Standard62HCCL3CDA
-
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Documents