Mouse Platelet-derived Growth Factor BB Recombinant

///Mouse Platelet-derived Growth Factor BB Recombinant

Mouse Platelet-derived Growth Factor BB Recombinant


accession P31240

Source Optimized DNA sequence encoding mouse PDGF-BB mature chain was expressed in Escherichia Coli.
Molecular weight MousePDGF-BB, generated by the proteolytic removal of the signal peptide and propeptide. The molecule has a calculated molecular mass of approximately 12 kDa. Recombinant PDGF-BB is a disulfide-linked homodimeric protein consisting of two 109 amino acid residue subunits, and migrates as an approximately 24 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC.
Biological Activity The ED(50) was determined by the dose-dependent proliferation of Balb/cT3 cells was found to be in the range of.0-2.0 ng/ml.

Endotoxin Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation Recombinant PDGF-BB was lyophilized from a 0.2 μm filtered solution in.5% glycine,.5% sucrose,.01% Tween80, mM Glutamic acid, pH.5.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
Storage The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

Molecular function Developmental-protein
Molecular function Growth-factor
Molecular function Mitogen


Efficient differentiation of intermediate lineages into vascular SMCs.

SMC differentiation medium CDM+PDGF-BB
  • The SMC subtypes, namely the neuroectoderm-derived SMC, lateral mesoderm-derived SMC, paraxial mesoderm-derived SMC are abbreviated as NE-SMC, LM-SMC and PM-SMC respectively.

ELISA of PDGF-BB secreted by EPCs in response to PDGFR-β transfection.

  • The concentration of PDGF-BB in the supernatant of culture medium was measured using ELISA.

Effect of calcitriol stimulation on VDR mRNA transcript, VDR protein expression, cell proliferation in PCASMCs.

±20 ng/ml PDGF-BB
  • Quiescent PCASMCs were stimulated with 20 ng/ml PDGF-BB with calcitriol for 24 h, and BrdU incorporation was analyzed .

Growth factor treatment and cell proliferation assay

  • Undifferentiated MVSCs and partially differentiated MVSCs (cultured in DMEM with 10% FBS for 3 weeks) were starved in DMEM with 1% FBS for 24 hours followed by the treatment of 10 ng/ml bFGF , 10 ng/ml PDGF-B or 10 ng/ml TGF-β1 for another 24 hours.
  • The cell proliferation was quantified by using Click-iT EdU Alexa Fluor 488 HCS Assay kit according to the instruction of the manufacturer.

Primary Culture of Epicardial Cells and Cardiomyocytes

  • Primary epicardial cells were cultured in 10% FBS/MEM containing recombinant soluble factors, such as TGFβ1 (10 ng/ml&systems), FGF2 (100 ng/mloche), , , , , PGF-BB (100 ng/ml) or retinoic acid (1 µmol/l), for 3 days.
  • The soluble factors were added on the day of heart removal.
  • The medium was replaced daily.
  • For western blot analysis, cells were treated with TGFβ1 for 2 days, beginning on the day following heart removal, at a final concentration of 1 ng/ml.

In vitro culture of stromal cells

  • Stromal cells were derived from E14.5 p190-B−/− and WT fetal livers.
  • The cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 20% fetal bovine serum (FBS, Omega Scientific, Tarzana, CA) and beta mercaptoethanol (2-ME , , ).
  • The adherent cells were grown to 70-80% confluence, passaged and sub-cultured in medium containing EGF (10ng/ml ) and PDGF (20ng/ml) .
  • The cells were depleted for CD11b using lineage depletion kit as per the manufacturer's protocol.
  • All experiments were performed on primary stroma of early passages (p3-p7) and exhibiting similar immunophenotype between the genotypes.

The effects of various glucose conditions on PDGF-mediated migration of retinal AC.

  • Transwell assay was performed with basal medium or medium containing PDGF-AA or PDGF-BB in lower compartment.

The effects of various glucose conditions on PDGF-mediated migration of retinal AC.

  • Transwell assay was performed with basal medium or medium containing PDGF-AA or PDGF-BB in lower compartment.

In vitro differentiation of Nes-GFP+ cells

  • LC differentiation For LC lineage differentiation, the Nes-GFP+ cells were replated in fresh differentiation-inducing medium containing phenol red-free DMEM/F12, 2% FCS, 10 ng/ml PDGF-BB , 1 ng/ml LH , 1 nM thyroid hormone , 70 ng/ml insulin-like growth factor 1 (IGF1), and ITS supplement , and they were incubated for 7 days, as previously described.
  • Differentiation was subsequently confirmed by RT-PCR and immunostaining for LC lineage markers (antibodies shown in