APPLICATION
Application Note
Secondary Lymphoid-tissue Chemokine (SLC) is a recently identified CC chemokine that is constitutively expressed in various lymphoid tissues and is a potent and specific chemoattractant for lymphocytes. Thus, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of SLC on the human T-lymphocyte leukemia cell line Jurkat. Briefly, Jurkat cells were seeded into the upper chambers (100 μl cell suspension, 1x10⁶ cells/ml in RPMI-1640 with FBS free) and SLC (1 ng/ml, 15 ng/ml and 150 ng/ml diluted separately in serum free RPMI-1640) was added in lower chamber with a polycarbonate filter (8um pore size) used to separate the two compartments. After incubation at 37ºC with 5% CO₂ for 1h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×100) and the number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter). The migrated Jurkat cells in low chamber at low magnification (×100) randomly, and the migrated cells were counted at high magnification (×400). SLC is able to induce migration of Jurkat cells, and the optimum chemotaxis of SLC occurs at 15-150 ng/ml.
Observed MW
18 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Ser24~Pro134 (NP_002980.1)
Expression System
E. coli
Purity
> 97%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
C-C motif chemokine ligand 21 , 6Ckine , CKb9 , ECL , SCYA21 , SLC , TCA4
Background
This antimicrobial gene is one of several CC cytokine genes clustered on the p-arm of chromosome 9. Cytokines are a family of secreted proteins involved in immunoregulatory and inflammatory processes. The CC cytokines are proteins characterized by two adjacent cysteines. Similar to other chemokines the protein encoded by this gene inhibits hemopoiesis and stimulates chemotaxis. This protein is chemotactic in vitro for thymocytes and activated T cells, but not for B cells, macrophages, or neutrophils. The cytokine encoded by this gene may also play a role in mediating homing of lymphocytes to secondary lymphoid organs. It is a high affinity functional ligand for chemokine receptor 7 that is expressed on T and B lymphocytes and a known receptor for another member of the cytokine family (small inducible cytokine A19). [provided by RefSeq, Sep 2014]
Database
Research Area
DATA IMAGES
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GTX00072-pro Functional Assay Image
The chemotactic effect of GTX00072-pro Human CCL21 protein (active). (A) Jurkat cells were seeded into the upper chambers and serum free RPMI-1640 without SLC was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 1hr. (B) Jurkat cells were seeded into the upper chambers and serum free RPMI-1640 with 150 ng/ml SLC was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 1hr.
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GTX00072-pro Image
SDS-PAGE analysis of GTX00072-pro Human CCL21 protein (active).
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GTX00072-pro Image
WB analysis of GTX00072-pro Human CCL21 protein (active).
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REFERENCE
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REVIEW
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