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Human Glial growth factor (GGF2) Recombinant

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$260.00$4,800.00

SKU: RKQ022979 Tags: , , , , , , , , , , ,

Description

Accession
Q02297
Source
Optimized DNA sequence encoding Human Neuregulin-1 isoform 9, GGF2 was expressed in HEK cells.
Molecular weight
Recombinant human GGF2 (Glial growth factor) is a monomeric protein consisting of 422 amino acid residue subunits, and migrates as an approximately 66 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-dependentphosphorylation of human MCF-7 cells was found to be <20ng/ml, corresponding to a specific activity ofx Units/mg.
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 GGF2 was lyophilized from a.2 μm filtered0.02M PB, 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
Molecular function

Methods

Culture and Differentiation of Human PSCs

  • Human ESCs [H9 (WA09, P35–45), were obtained from WiCell Inc., Madison, WI, ] 5 cells/cm2 in medium'>medium'>N2B27 medium'>medium or 1× N2, 0.5× B27 and 0.5× G21 supplement (referred to as medium'>NBG medium'>medium) plus 20 ng/ml of basic FGF.
  • The PSA-NCAM-negative cells were also cultured in the same condition as the culture of PSA-NCAM-positive other than the density of 1×105 cells/cm2.
  • Culture medium was changed every day and cells were passaged every 2–3 days.
  • If cells did not display sufficient purity, an additional round of cell sorting was performed to enhance the homogeneity of hNPCPSA-NCAM+.
  • For further differentiation, hNPCPSA-NCAM+ were seeded at a low density (>2×104 cells/cm2) and cultured in differentiation media for 3 weeks.
  • Differentiation media was composed of N2B27 or NBG medium supplemented with 0.1% fetal bovine serum and 200 µM of ascorbic acid .
  • Media changes were…

Generation of human neurons derived from hPSC

  • hiPSCs were generated from fibroblasts from SPG11 patients (SPG11-1 and SPG11-2) and control subjects (CTRL-1 and CTRL-2) as previously described (TM Express).
  • Terminal medium'>differentiation of NPCs towards neuronal cells was initiated in medium'>neural medium'>differentiation medium [NDM: NIM supplemented with 20 ng/ml brain-derived neurotrophic factor , 20 ng/ml glial cell line-derived neurotrophic factor , 1 mm dibutyryl-cyclic AMP and 200 nm ascorbic acid ] at a density of 40 000 cells/cm2 on PORN/laminin-coated plates or glass coverslips.
  • Neuronal cultures were kept for differentiation under these conditions from 12 to 40 days with a half medium change every week.
  • Since we have used one NPC/neuronal line from each control and SPG11 patient groups, for the sake of simplicity for the readers, we will hereafter refer to the neuronal lines as CTRL-1, CTRL-2, SPG11-1 and SPG11-2 from the control and SPG11 patients, respectively.

Dopaminergic neuronal induction of bone marrow-derived mesenchymal stem cells

  • Passage 3 bone marrow-derived mesenchymal stem cells were trypsinized and subcultured onto polylysine-coated coverslips in multiwell plates.
  • Each plate contained 1 × 105 cells.
  • At approximately 80% confluence, bone marrow-derived mesenchymal stem cells were induced into dopaminergic neurons using different inducers as follows: in the Xiangdan injection group, cells were pre-induced in L-Dulbecco's-modified Eagle's medium containing 10% fetal bovine serum and 25 ng/mL basic fibroblast growth factor for 24 hours, followed by L-Dulbecco's-modified Eagle's medium and 20% Xiangdan injection (containing 1 000 g/L salvia miltiorrhiza, 1 000 g/L dalbergia odorifera (No. Z13021387), Hebei Tiancheng Pharmaceutical Co., Ltd., China) for 3 hours; in the all-trans retinoic acid + glial cell line-derived neurotrophic factor group, cells were induced in medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>neurobasal cells.
  • At approximately 80% confluence, bone marrow-derived mesenchymal stem cells were induced into dopaminergic neurons using different inducers as follows: in the Xiangdan injection group, cells were pre-induced in L-Dulbecco's-modified Eagle's medium containing 10% fetal bovine serum and 25 ng/mL basic fibroblast growth factor for 24 hours, followed by L-Dulbecco's-modified Eagle's medium and 20% Xiangdan injection (containing 1 000 g/L salvia miltiorrhiza, 1 000 g/L dalbergia odorifera (No. Z13021387), Hebei Tiancheng Pharmaceutical Co., Ltd., China) for 3 hours; in the all-trans retinoic acid + glial cell line-derived neurotrophic factor group, cells were induced in medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>neurobasal medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium containing 25 ng/mL basic fibroblast growth factor and 2% B27 for 24 hours, followed by medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>neurobasal medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium containing 2% B27, 1 μM all-trans retinoic acid and 50 ng/mL glial cell line-derived neurotrophic factor for 6 days; in the sonic hedgehog + fibroblast growth factor 8 group, cells were induced in medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>neurobasal medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium containing 25 ng/mL basic fibroblast growth factor and 2% B27 for 24 hours, followed by medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium'>neurobasal medium'>medium'>medium'>medium'>medium'>medium'>medium'>medium containing 2% B27, 250 ng/mL sonic hedgehog and 100 ng/mL fibroblast growth factor 8 for 12 days; in the control group, cells were cultured without any inducers.

Neural differentiation

  • iPSC-derived neural progenitors were generated by either embryoid body (EB)-based or monolayer differentiation according to established protocols [
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