Human Platelet Derived Growth Factor-AA Recombinant

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Human Platelet Derived Growth Factor-AA Recombinant

$70.00$3,500.00


accession P04085


Source Optimized DNA sequence encoding Human PDGF-AA mature chain was expressed in CHO Cells.
Molecular weight Human PDGF-AA, is generated by the proteolytic removal of the signal peptide and propeptide. The molecule has a calculated molecular mass of approximately kDa. Recombinant PDGF-AA is a disulfide-linked homodimeric protein consisting of two 125 amino acid residue subunits, migrates due to glycosylation as an approximately 29 kDa protein under non-reducing and as 14 kDa under reducing conditions in SDS-PAGE.
Purity >98%, as determined by SDS-PAGE and HPLC
Biological Activity The ED(50) was determinedby the dose-dependent stimulation of thymidine uptake by Balb/cT3 cells is ≤ ng/ml, corresponding to a specific activity of > x units/mg
Protein Sequence MRTLACLLLL GCGYLAHVLA EEAEIPREVI ERLARSQIHS IRDLQRLLEI DSVGSEDSLD TSLRAHGVHA TKHVPEKRPL PIRRKRSIEE AVPAVCKTRT VIYEIPRSQV DPTSANFLIW PPCVEVKRCT GCCNTSSVKC QPSRVHHRSV KVAKVEYVRK KPKLKEVQVR LEEHLECACA TTSLNPDYRE EDTGRPRESG KKRKRKRLKP T
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-AAwas lyophilized from a.2 μm filteredmM NaCl,mM Tris solution pH7.0.
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.


Interactor P01730 CD4_HUMAN
Interactor P02751 FINC_HUMAN
Interactor P16234 PGFRA_HUMAN
Molecular function Developmental-protein
Molecular function Growth-factor
Molecular function Mitogen

Methods

Differentiation of spinal cord NSPC

  • NSPC were assessed for multipotentiality by plating onto matrigel in SFM with 1% fetal bovine serum (FBS) in the absence of growth factors for 4 weeks to allow the cells to differentiate.
  • To assess the effect of exogenous factors on NSPC differentiation, NSPC were plated in EGF/FGF2 medium at a density of 105 cells/well into 24-well culture plates coated with matrigel.
  • Cultures were incubated for 1 week, and then the medium was replaced with one of the following factors in SFM: 40 and 100 ng/ml PDGF-AA to promote oligodendrocyte differentiation, and 1 and 4 mM dbcAMP to promote neuronal differentiation.
  • Controls included 1% FBS and SFM.
  • Cultures were incubated at 37°C for an additional 4 weeks and the media changed every week.
  • The phenotypic expression pattern of NSPC in differentiating culture conditions was examined by immunostaining as described below.
  • The total number of cells counted ranged from 180 – 400 cells…
  • NSPC were assessed for multipotentiality by plating onto matrigel in SFM with 1% fetal bovine serum (FBS) in the absence of growth factors for 4 weeks to allow the cells to differentiate.
  • To assess the effect of exogenous factors on NSPC differentiation, NSPC were plated in EGF/FGF2 medium at a density of 105 cells/well into 24-well culture plates coated with matrigel.
  • Cultures were incubated for 1 week, and then the medium was replaced with one of the following factors in SFM: 40 and 100 ng/ml PDGF-AA to promote oligodendrocyte differentiation, and 1 and 4 mM dbcAMP to promote neuronal differentiation.
  • Controls included 1% FBS and SFM.
  • Cultures were incubated at 37°C for an additional 4 weeks and the media changed every week.
  • The phenotypic expression pattern of NSPC in differentiating culture conditions was examined by immunostaining as described below.
  • The total number of cells counted ranged from 180 – 400 cells per marker.

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