Buffer Formulation	20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300.
Traits	Freeze-dried powder
Purity	> 95%
Isoelectric Point	4.8
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

MIP-1a (macrophage inflammatory protein 1-alpha) also known as Chemokine (C-C motief) ligand 3 (CCL3), is a cytokine belonging to the CC chemokine family that is involved in the recruitment and activation of macrophages, monocytes and neutrophils. In this case, chemotaxis assay used 24-well microchemotaxis system was undertaken to evaluate the chemotactic effect of MIP-1a on the human monocytic cell line THP1. Briefly, THP1 cells were seeded into the upper chambers (100µl cell suspension, 106 cells/ml in RPMI 1640 with 0.5% FBS) and MIP-1a (100ng/mL, diluted 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 5h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×40) and the number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter).
By counting migrated cells in low chamber at high magnification (×400) randomly, it was shown that a mean of 41.2 THP1 cells/field migrated towards serum free RPMI 1640 medium with 100ng/mL MIP-1a, while only 3.6 THP1 cells/field migrated towards serum free RPMI 1640 medium. And the migrated THP1 cells in low chamber at low magnification (×40) was shown in Figure 1.

 

USAGE

 

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

 

STORAGE

 

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

 

STABILITY

 

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

 

Image


SDS-PAGE Image

 

Figure. Western Blot; Sample: APA092Hu61; Antibody: PAA092Hu06.

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