/, TGF-beta family, Recombinant Human Cytokines/Human Bone Morphogenetic protein-7 Recombinant

Human Bone Morphogenetic protein-7 Recombinant

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$70.00$3,500.00

SKU: RKP180751 Tags: , , , , , ,

Description

Methods

rBMP7 rescues Ngn2 mRNA expression in cultured E14.5 Bmp7-deficient cortical cells.

  • RT-PCR analysis of serial 4-fold cDNA dilutions (1, ¼, 1/16, 1/64, 1/256 cDNA input) for Ngn2 and Pax6 in isolated E14.5 wild-type (wt) or Bmp7

Cell culture and immunoblotting

  • Cranial fibroblasts were isolated from E14.5 C57BL/6 mouse embryos.
  • The mid-facial region was dissected from embryo heads and incubated in 0.05% trypsin-EDTA solution for 1 h at 37°C, after which the tissue was triturated by repeated aspiration.
  • Cells were centrifuged, resuspended in Dulbecco's Minimal Essential Medium (DMEM;) containing 10% fetal calf serum (FCS), plated onto 10 cm cell culture dishes, and incubated at 37°C with 6% CO2.
  • At confluency, cranial fibroblasts were passaged by trypsinization.
  • Bmp7 stimulation experiments were performed in 24-well dishes with cells from the second passage.
  • Fibroblasts were first starved for 24 h in DMEM/0.3% FCS, before the medium was changed to DMEM/0.3% FCS containing human recombinant Bmp7 (, , 0, 10, 20, 50, and 100 ng/ml, respectively).
  • For positive controls, wells were incubated with DMEM/10% FCS.
  • Cell supernatant was collected from individual wells after 24 h, run on…

Eggs and Embryo Manipulations

  • Fertilized White Leghorn chicken eggs were incubated at 38°C and the embryos staged according to Hamburger and in situ hybridization.

Specification of intermediate mesoderm (IM) from the hESC-derived PS cells.

  • (b) Comparison of OSR1 transcripts by combinatorial treatments with exogenous growth factors of retinoic acid , BMP7 (B7) and FGF2 (F2).
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