Application Note
PDGFA (Platelet-derived growth factor subunit A) is a Growth factor that plays an essential role in the regulation of embryonic development, cell proliferation, cell migration, survival and chemotaxis. PDGFA has been described as a chemoattractant for monocytes and proven to be able to induce chemotactic migration of THP-1 cells. Therefore, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of PDGFA on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100 μl cell suspension, 1x10⁶cells/ml in RPMI-1640 with 0. 5% FBS) and PDGFA (1. 5 ng/ml, 7. 5 ng/ml and 15 ng/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 (×40) and the number of migrated cells were counted at high magnification (×400) randomly. PDGFA is able to induce migration of THP-1 cells, and the optimum chemotaxis of PDGFA occurs at 1. 5 ng/ml.
Observed MW
17 kDa.
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; Ala94~Asp194 (NP_032834.1)
Expression System
E. coli
Purity
> 95%
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.