Human IGF-I Recombinant
Categories: IGF familyIGF familyRecombinant Human Cytokines$70.00 – $220.00
Description
Accession
P01343
Source
Optimized DNA sequence encoding Human Insulin Growth Factor -1mature chain was expressed in Escherichia Coli.
Molecular weight
Native Human Insulin Growth Factor-1 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 8 kDa. Recombinant Human IGF-1 is a monomeric protein consisting of 71 amino acid residue subunits. IGF-1 migrates as an approximately 8 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED50 was determined by a cell proliferation assay using FDC-P1 cells is ≤1.0 ng/ml, corresponding to a specific activity of ≥1 x107 units/mg.
Protein Sequence
MGKISSLPTQ LFKCCFCDFL KVKMHTMSSS HLFYLALCLL TFTSSATAGP ETLCGAELVD ALQFVCGDRG FYFNKPTGYG SSSRRAPQTG IVDECCFRSC DLRRLEMYCA PLKPAKSA RS VRAQRHTDMP KTQKEVHLKN ASRGSAGNKN YRM
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
Recombinant Human IGF1was lyophilized from a 0.2 μm filtered PBS solution, pH.4.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 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 at2° -8° 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.
Methods
Effect of vorinostat on IGF-I-mediated signal transduction.
- Ishikawa and USPC-2 cells were treated for 24 h with vorinostat (or left untreated) and/or IGF-I during the last 10 min of the incubation period.
Induction of lamellipodia formation and invasion by IGF-I in MDA-MB-231 cells but not MCF7 cells.
- (a) After stimulation with or without (mock) IGF-I, cells were stained with phalloidin.
Cell preparation
- Human ESC-CMs and non-cardiac hESC derivatives were harvested and cryopreserved after 16–18 days under differentiating conditions.
- One day prior to harvest, cells were subjected to a pro-survival protocol, previously shown to enhance engraftment post-transplantation.
- In brief, cultures were heat-shocked with a 30-minute exposure to 43 °C medium, followed by RPMI-B27 medium supplemented with IGF-1 (100 ng/ml) and cyclosporine A (0.2 μM, , ).
- One day later, cultures were harvested with 0.25% trypsin/0.5 mM EDTA and cryopreserved as previously described.
Materials
-
Hank's Balanced Sodium Salt (HBSS), medium'>Dulbecco's medium'>modified medium'>Eagle's medium (DMEM) high glucose (4.5 g/L), alpha minimum essential medium, phosphate-buffered salt (PBS), penicillin/streptomycin, trypsin/EDTA (0.05%/0.53 mM),
L -glutamine, superscript III kit, NuPAGE 4%–12% Bis–Tris gel, bone morphogenetic protein-2 (BMP-2), and polyvinylidene difluoride (PVDF)membranes were obtained from . - Low-molecular weight heparin (4,500 g/mol) was obtained from (,
http://www.sanofi.com/ ). - 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) reagents were from (,
http://www.promega.com/products ). - Total collagens and GAG quantification kits were from (,
http://www.biocolor.co.uk ). - Brilliant SYBR Green Master Mix was obtained from ( , The
http://www.stratagene.com ). - PCR primers were synthesized by MWG (,
http://www.mwg-biotech.com ). - TGF-β1 and Insulin-like growth factor-1 (IGF-1) were obtained from (,
http://www.peprotechec.com ). - Protein content was determined using the assay (, ,
http://www.piercenet.com ). - The Western blot detection system was obtained from GE Healthcare (Buckin-ghamshire, UK,
http://www3.gehealthcare.com ). - All other chemicals were obtained from standard laboratory suppliers and were of the highest purity…
Effects of IGF-1-containing gelatin hydrogel on neurogenesis in the SVZ.
- (b) Coronal sections of brains that received an injection of microspheres containing IGF-1 or microspheres plus PBS, showing DCX+ new neurons in the SVZ (green) (n = 5 animals for each group).
Effects of Activin A on paraxial mesodermal differentiation of mouse iPS cells.
- The cultures also contained BMP4 (10 ng/ml), IGF-1 (10 ng/ml), LiCl (5 mM), and Shh (10 ng/ml).
In vitro Embryo Cultures
- Tri-pronuclear zygotes were cultured individually in 30 µl microdrops containing the Global-Medium with 5% human serum albumin in the presence or absence of EGF, IGF-I, GM-CSF, BDNF, and CSF-1 , all at 10 ng/ml.
- The culture medium was renewed every 48 h. Embryonic development was evaluated at 96, 120, and 144 h after culture.
In vitro Embryo Cultures
- Normally fertilized embryos frozen on day 3 of culture by slow cooling were thawed by using a 2-step thawing protocol 2, 5% O2 and 90% N2 with or without growth factor mixtures containing 10 ng/ml of EGF, IGF-I, GM-CSF, BNF, CSF-1, artemin, and GNF .
- Individual embryos were cultured for 72 h in a 30 µl drop of medium and their development was evaluated.
- The doses of these growth factors chosen for these experiments were based on previous studies (BDNF
Cell cycle analysis of MCF-7 cells in the presence of IGF-1 at 8 h. MCF-7 cells were serum starved for 24 h. At 22 h after serum starvation the cells were treated with 10 μM auraptene in 0.01% DMSO.
- At 24 h serum starvation, the cells were treated with IGF-1 (10 ng/mL).
Effect of metformin on IGF-I-mediated signal transduction and mTOR and Ampk signalling pathway in endometrial cancer cells.
- A, Ishikawa, ECC-1, USPC-2 and USPC-1 cells were treated with metformin (10 mM) for 24 h (or left untreated) in the presence or absence of IGF-I (50 ng/ml) during the last 10 min of the incubation period.
