Difference between revisions of "Part:BBa K3562013"
 
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The protocols of our experiments are shown in our Notebook part of our wiki. Links are shown below:
 
The protocols of our experiments are shown in our Notebook part of our wiki. Links are shown below:
 
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[https://2020.igem.org/wiki/images/a/a6/T--WHU-China--protocol_11_12.pdf]
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[https://2020.igem.org/wiki/images/a/a6/T--WHU-China--protocol_11_12.pdf Protocols of transwell experiments]
 
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[https://2020.igem.org/wiki/images/3/36/T--WHU-China--14.pdf]
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[https://2020.igem.org/wiki/images/3/36/T--WHU-China--14.pdf Protocols of flow cytometry]
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===Reference===
 
===Reference===
 
[1][https://www.ebi.ac.uk/QuickGO/term/GO:0008009 Chemokine activity definition]
 
[1][https://www.ebi.ac.uk/QuickGO/term/GO:0008009 Chemokine activity definition]
 
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[2][https://www.uniprot.org/uniprot/P13500 UniProtKB - P13500 (CCL2_HUMAN)]
 
[2][https://www.uniprot.org/uniprot/P13500 UniProtKB - P13500 (CCL2_HUMAN)]

Latest revision as of 15:33, 26 October 2020


CCL2

A ligand for C-C chemokine receptor CCR2 that exhibits a chemotactic activity for monocytes and basophils but not neutrophils or eosinophils.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

Chemokines are a family of small chemotactic cytokines, and the name is derived from their ability to induce directed chemotaxis in nearby responsive cells. All chemokines possess a number of conserved cysteine residues involved in intramolecular disulfide bond formation. Some chemokines can be induced during an immune response to recruit cells of the immune system to a site of infection, while others are considered homeostatic and are involved in controlling the migration of cells during normal processes of tissue maintenance or development. Chemokines are found in all vertebrates, some viruses and some bacteria.

CCL2 acts as a ligand for C-C chemokine receptor CCR2. CCL2 signals through binding and activation of CCR2 and induces a strong chemotactic response and mobilization of intracellular calcium ions. It exhibits a chemotactic activity for monocytes and basophils but not neutrophils or eosinophils. CCL2 may be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis.

Experiments and Results

We used the Transwell experiment(chamber diameter: 6.5 mm, Pore diameter of PC membrane: 8 μm, 48 pores/board) combined with flow cytometry to verify the chemotactic ability of this chemokine on the THP1 cell line.(Fig.1)

Figure 1 Chemotaxis of CCL2 to THP1(shown by cell number). Chemokine concentration is 10 nmol/l. (n=3)

Besides, we also tested the activity of six other chemokines.(Fig.2)

Figure 2 Chemotaxis index(CI) of 7 chemokines to THP1.(n=3)

The protocols of our experiments are shown in our Notebook part of our wiki. Links are shown below:

Protocols of transwell experiments
Protocols of flow cytometry

Reference

[1]Chemokine activity definition
[2]UniProtKB - P13500 (CCL2_HUMAN)