Human Bone Morphogenetic protein-7 Recombinant
Categories: Recombinant Human CytokinesTGF-beta familyTGF-beta family$70.00 – $3,500.00
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.