////Mouse Hepatocyte Growth Factor Recombinant

Mouse Hepatocyte Growth Factor Recombinant

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$160.00$2,700.00

SKU: RKQ08048 Tags: , , ,

Description

Accession
Q08048
Source
Optimized DNA sequence encoding Mouse Hepatocyte Growth Factor mature chain was expressed in Chinese Hamster Ovary cell line.
Molecular weight
Native human Hepatocyte Growth Factor is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 80kDa. Recombinant HGF is a glycosylated disulfide-linked homodimeric protein consisting ofalpha chain (463)and the beta chain (233)amino acid residue subunits, and migrates as an approximately 75 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the dose-dependent stimulation of the proliferation ofmurine BNL-CL2cell li was found to be in the range of 20.0-40.0 ng/ml.

Protein Sequence
MMWGTKLLPV LLLQHVLLHL LLLHVAIPYA EGQKKRRNTL HEFKKSAKTT 50 LTKEDPLLKI KTKKVNSADE CANRCIRNRG FTFTCKAFVF DKSRKRCYWY 100 PFNSMSSGVK KGFGHEFDLY ENKDYIRNCI IGKGGSYKGT VSITKSGIKC 150 QPWNSMIPHE HSFLPSSYRG KDLQENYCRN PRGEEGGPWC FTSNPEVRYE 200 VCDIPQCSEV ECMTCNGESY RGPMDHTESG KTCQRWDQQT PHRHKFLPER 250 YPDKGFDDNY CRNPDGKPRP WCYTLDPDTP WEYCAIKTCA HSAVNETDVP 300 METTECIQGQ GEGYRGTSNT IWNGIPCQRW DSQYPHKHDI TPENFKCKDL 350 RENYCRNPDG AESPWCFTTD PNIRVGYCSQ IPKCDVSSGQ DCYRGNGKNY 400 MGNLSKTRSG LTCSMWDKNM EDLHRHIFWE PDASKLNKNY CRNPDDDAHG 450 PWCYTGNPLI PWDYCPISRC EGDTTPTIVN LDHPVISCAK TKQLR>VVNGI 500 PTQTTVGWMV SLKYRNKHIC GGSLIKESWV LTARQCFPAR NKDLKDYEAW 550 LGIHDVHERG EEKRKQILNI SQLVYGPEGS DLVLLKLARP AILDNFVSTI 600 DLPSYGCTIP EKTTCSIYGW GYTGLINADG LLRVAHLYIM GNEKCSQHHQ 650 GKVTLNESEL CAGAEKIGSG PCEGDYGGPL ICEQHKMRMV LGVIVPGRGC 700 AIPNRPGIFV RVAYYAKWIH KVILTYKL
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
Mouse HGF was lyophilized from a 0.2 μm filtered solution in 2.5% glycine, 0.5% sucrose, 0.01% Tween80, 5 mM Glutamic acid, pH 4.5.
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 2 years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at 2° - 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.
Interactor
Interactor
P46414
Molecular function
Molecular function

Methods

Met knockdown reduces BxPC-3 and ASPC-1 cell survival.

  • BxPC-3 or ASPC-1 cells infected with recombinant lentivirus expressing Met knockdown shRNAs (2 or 5) or a non targeting shRNA were treated without or with HGF and examined by Western analysis for pMet (Y1234/1235), Met, pErk1/2, Erk1/2, pAKT, AKT and β-actin levels (n = 3).
  • Hepatic. Cells were seeded at a concentration of 5,000 cells/cm2 on tissue culture plastic plates and coverslips coated with Matrigel and cultured in high glucose DMEM supplemented with 1% Penicillin/Streptomycin , 2 mmol/l L-Glutamine and 10% FBS for 3 days.
  • The media were then changed to high glucose DMEM supplemented with 15% FBS, 1% Penicillin/Streptomycin, 2 mmol/l L-Glutamine, 300 µmol/l Monothioglycerol , 20 ng/ml Hepatocyte Growth Factor , 10 ng/ml Oncostatin M , 10−7 Dexamethasone , 100 ng/ml FGF4 , and 1X ITS .
  • The cells were allowed to differentiate for 21 days and then fixed and stored in PBS for immunofluorescence.
  • The differentiation media were collected and analyzed for the presence of urea secreted by the differentiated cells.
  • Urea was subsequently measured using the Urea/Ammonia determination kit (R-Biopharm AG, Darmstadt, Germany) according to the manufacturers' instructions.
  • Cells were subsquently assessed for expression of the hepatocyte markers ALBUMIN and…

2.4. In Vitro Hepatic Differentiation

  • Prior to starting the hepatic differentiation, medium'>MSCs at passage 5 were maintained in the regular medium'>MSC culture medium until at 80–90% confluence.
  • The hepatic differentiation was elicited in the differentiation-inducing medium, which consisted of MEM supplemented with 10% FBS, 10 ng/mL hepatocyte growth factor (HGF) , 10 ng/mL basic fibroblast growth factor (bFGF) and 10 ng/mL oncostatin M (&system).
  • The medium was changed every 3 days, and the cells were cultured for 8 days.

In vitro differentiation of hepatocyte-like cells from undifferentiated iPS cells

  • >Stage 3: Differentiation of Premature Hepatocyte-Like Cells (5–11).
  • At day 5, the medium was replaced by Knockout DMEM supplemented with 1% KSR, 1% nonessential amino acids, 1% l-glutamic acid, 1% dimethyl sulfoxide (DMSO) , and 100 ng/ml hepatocyte growth factor (HGF) to induce premature hepatocyte-like cells.
  • The medium was replaced every two days.

Cell Culture

  • NPCs were prepared from striatum of embryonic day 17 to 18 (E17–18) old rats and the cells were plated in dishes (5×106 cells per 10 cm dish , , ) and cultivated in a medium'>serum-free medium'>DMEM/F12 medium in the presence of B27 supplement and 20 ng/ml epidermal growth factor (EGF) at +37°C in 5% CO2

Conditioned media from BMPC regulates miR-155 expression and fibrogenic response in mouse cardiac fibroblasts in vitro.

  • Neutralizing antibodies against HGF (HGF-Ab) reversed this effect as compared to IgG treated cells.

Cell culture and hepatic cell differentiation

  • Huh7 and Huh7.5 cells were provided by Jane C. Moores (The Reagent of the University of California, Oakland, CA).
  • The cells were maintained in medium'>Dulbecco's medium'>modified medium'>Eagle's medium (DMEM;, , ) containing 10% FBS and 1% penicillin/streptomycin in a humidified incubator at 37°C with 5% CO2.
  • For hepatic differentiation, passages 3–7 of AT-hMSCs were plated at 1×104 cells/cm2 and incubated overnight.
  • The cells were then washed with phosphate-buffered saline (PBS) and incubated in basal medium [60% DMEM-low glucose , 40% MCDB201 , and 1% penicillin/streptomycin] with 20 ng/ml EGF and 10 ng/ml bFGF for 2 days.
  • After incubation, hepatogenic cytokines and growth factors were sequentially added as follows: days 0–7 (step 1), basal medium with 20 ng/ml HGF and 10 ng/ml bFGF; and days 7–21 (step 2), basal medium containing 20 ng/ml oncostatin M , 1 µmol/L dexamethasone…

Differentiation of hESCs into dopaminergic neurons, osteoblasts, and hepatocytes

  • Derivation of hepatocytes from hESCs was carried out as previously described (μM dexamethasone for 5 days.
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