Active Monocyte Chemotactic Protein 4 (MCP4)

单核细胞趋化蛋白4(MCP4)活性蛋白

[ PROPERTIES ]

Source: Eukaryotic expression. Host: Yeast

Residues: Phe17~Thr98

Tags: N-terminal His-tag

Purity: >95%

Endotoxin Level: <1.0EU per 1μg (determined by the LAL method). Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% sarcosyl, 5% trehalose, and Proclin300. Applications: Cell culture; Activity Assays; In vivo assays. (May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 10.0

Predicted Molecular Mass: 10.7kDa

Accurate Molecular Mass: 13kDa as determined by SDS-PAGE reducing conditions. 

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex. 

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles.

Store at 2-8ºC for one month.

Aliquot and store at -80ºC for 12 months.

Stability Test: The thermal stability is described by the loss rate. The loss ratewas determined by accelerated thermal degradation test, that is, incubate theprotein at 37ºC for 48h, and no obvious degradation and precipitation wereobserved.The loss rate is less than 5% within the expiration date underappropriate storage condition

[ SEQUENCE ]

[ ACTIVITY ]

CCL13 (C-C motif chemokine 13) is a chemotactic factor that attracts monocytes,lymphocytes, basophils and eosinophils, which belongs to the CC chemokine subfamily. It has been reported that CCL13 can induce chemotactic migration of THP-1 cells. Therefore, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of CCL13 on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100µL cell suspension,10 ⁶cells/mL in RPMI 1640 with 0.5% FBS) and CCL13 (50ng/mL and 100ng/mL diluted separately in serum free RPMI 1640) was added in lower chamber with a polycarbonate filter (8µm pore size) used to separate the two compartments. After incubation at 37ºC with 5% CO2 for 3h, 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). Result: CCL13 is able to induce migration of THP-1 cells. The migrated THP-1 cells in low chamber at low magnification (×100) were shown in Figure 1. Five fields of each chamber were randomly chosen to count the migrated cells at high magnification (×400) and the statistical data was shown in Figure 2.

[ IDENTIFICATION ]

Figure 3. Gene Sequencing (extract)