Human Glial growth factor (GGF2) Recombinant

HomeEGF family, Recombinant Human Cytokines

Human Glial growth factor (GGF2) Recombinant

$260.00 – $4,800.00

SKU: RKQ022979 Categories: EGF family, Recombinant Human Cytokines Tags: ARIA, GGF, Glial growth factor, heregulin, HGL, hrg, HRGA, NDF, nrg1, Pro-NRG1, Q02297, SMDF

Description

Methods (4)


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	P21860 ERBB3_HUMAN
Molecular function	Growth-factor

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 hNPC^PSA-NCAM+. For further differentiation, hNPC^PSA-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… 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 hNPC^PSA-NCAM+. For further differentiation, hNPC^PSA-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 performed every two days. For differentiation to midbrain dopaminergic (A) neurons and spinal motor neurons, hNPC^PSA-NCAM+ were treated with 200 ng/ml Shh and 100 ng/ml FGF8 or 100 ng/ml Shh and 0.5 µM retinoic acid (A) for 7 or 8 days, respectively. The cells were re-plated on new Matrigel-coated dishes at a low density and cultured in differentiation media supplemented with 10 ng/ml brain-derived neurotrophic factor (BDNF) , and 10 ng/ml glial cell-derived neurotrophic factor (GDNF) for 3 weeks. Read more
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. Read more
Neural differentiation iPSC-derived neural progenitors were generated by either embryoid body (EB)-based or monolayer differentiation according to established protocols [