Human Hepatocyte Growth Factor Recombinant

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Human Hepatocyte Growth Factor Recombinant

$70.00$4,300.00


accession P14210


Source Optimized DNA sequence encoding Human Hepatocyte Growth Factor mature chain was expressed in Chinese Hamster Ovary cell line.
Molecular weight Native humanHepatocyte Growth Factoris generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximatelykDa. Recombinant HGF is a glycosylated disulfide-linked homodimeric protein consisting ofalpha chain (463)and the beta chain (234)amino acid residue subunits, and migrates as an approximately 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.0-40.0 ng/ml.

Protein Sequence MWVTKLLPAL LLQHVLLHLL LLPIAIPYAE GQRKRRNTIH EFKKSAKTTL IKIDPALKIK TKKVNTADQC ANRCTRNKGL PFTCKAFVFD KARKQCLWFP FNSMSSGVKK EFGHEFDLYE NKDYIRNCII GKGRSYKGTV SITKSGIKCQ PWSSMIPHEH SFLPSSYRGK DLQENYCRNP RGEEGGPWCF TSNPEVRYEV CDIPQCSEVE CMTCNGESYR GLMDHTESGK ICQRWDHQTP HRHKFLPERY PDKGFDDNYC RNPDGQPRPW CYTLDPHTRW EYCAIKTCAD NTMNDTDVPL ETTECIQGQG EGYRGTVNTI WNGIPCQRWD SQYPHEHDMT PENFKCKDLR ENYCRNPDGS ESPWCFTTDP NIRVGYCSQI PNCDMSHGQD CYRGNGKNYM GNLSQTRSGL TCSMWDKNME DLHRHIFWEP DASKLNENYC RNPDDDAHGP WCYTGNPLIP WDYCPISRCE GDTTPTIVNL DHPVISCAKT KQLRVVNGIP TRTNIGWMVS LRYRNKHICG GSLIKESWVL TARQCFPSRD LKDYEAWLGI HDVHGRGDEK CKQVLNVSQL VYGPEGSDLV LMKLARPAVL DDFVSTIDLP NYGCTIPEKT SCSVYGWGYT GLINYDGLLR VAHLYIMGNE KCSQHHRGKV TLNESEICAG AEKIGSGPCE GDYGGPLVCE QHKMRMVLGV IVPGRGCAIP NRPGIFVRVA YYAKWIHKII LTYKVPQS
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 Interleukin-6 was lyophilized from a.2 μm filtered solution in.5% glycine,.5% sucrose,.01% Tween80, mM Glutamic acid, pH.5.
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 P08581 MET_HUMAN
Interactor P01730 CD4_HUMAN
Interactor Q04756 HGFA_HUMAN
Molecular function Growth-factor
Molecular function Serine-protease-homolog

Methods

Silencing of mda-9/syntenin in 92.1 uveal melanoma cells inhibits in vitro invasion and HGF mediated signaling.

recombinant HGF
  • B: Invasion of matrigel membranes by 92.1 cells towards different stimuli: medium with 10% serum , 50% conditioned medium from MG63 cell line , 100 ng/ml recombinant HGF in 0.1% serum.

Hepatogenic differentiation

  • Cells were seeded at a concentration of 5000 cells/cm2 on tissue culture plastic plates and coverslips coated with Matrigel and cultured in high glucose DMEM supplemented with 1% penicillin/streptomycin , 2 mM 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 mM L-glutamine, 300 µM monothioglycerol , 20 ng/ml hepatocyte growth factor , 10 ng/ml oncostatin M , 10−7 dexamethasone , 100 ng/ml FGF4 and 1X ITS (insulin, transferrin, selenium).
  • The cells were allowed to differentiate for 21 days and then fixed and stored in PBS for immunofluorescence.
  • The differentiation media were collected and analysed 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, with positive control being urea (as per manufacturer's instructions) and…
  • Cells were seeded at a concentration of 5000 cells/cm2 on tissue culture plastic plates and coverslips coated with Matrigel and cultured in high glucose DMEM supplemented with 1% penicillin/streptomycin , 2 mM 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 mM L-glutamine, 300 µM monothioglycerol , 20 ng/ml hepatocyte growth factor , 10 ng/ml oncostatin M , 10−7 dexamethasone , 100 ng/ml FGF4 and 1X ITS (insulin, transferrin, selenium).
  • The cells were allowed to differentiate for 21 days and then fixed and stored in PBS for immunofluorescence.
  • The differentiation media were collected and analysed 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, with positive control being urea (as per manufacturer's instructions) and negative control undifferentiated e-CSC and l-CSC.
  • Cells were subsquently assessed for expression of the hepatocyte markers ALBUMIN and ALPHA FETO-PROTEIN (AFP) using confocal microscopy.
  • Positive control were HepG2 cells (gift from Wei Cui) and negative controls were undifferentiated e-CSC and l-CSC cells.

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Effects of HGF-containing gelatin hydrogel on the number of new neurons migrating in the ischemic striatum after middle cerebral artery occlusion.

recombinant human HGF
  • (c) Coronal sections stained for DCX (green) from brains that received a PBS or HGF injection with (n = 6 animals for each group) or without (n = 4 animals for each group) gelatin hydrogel.

HGF up-regulates the expression of HLX1 in HTR-8/SVneo cells.

HGF
  • HTR-8/SVneo cells were treated with the indicated concentrations of HGF for 48 h. The steady-state mRNA level of HLX1 was examined by qPCR and normalized to a β-actin internal control.

Isolation of primary human LSECs

  • Human liver endothelial cells were isolated from liver tissue from patients undergoing surgical liver resection.
  • The tissue was collected with informed consent and under local ethics committee approval of the University of Bern.
  • Approximately 30 g of liver was cut into 2 mm3 pieces and digested in 50 ml PBS containing 10 mg/ml collagenase 1A for 90 min at 37 °C.
  • The resultant solution was then filtered through a sterile 0.5 mm steel mesh, the supernatant centrifuged at 300 × g for 8 min, and the pellet then resuspended on 10 ml PBS.
  • This digest was then layered on a 33%/77% Percoll density gradient, centrifuged for 30 min at 800 × g 4 °C and the non-parenchymal layer removed, pelleted and resuspended in 500 μl of PBS.
  • Endothelial cells were isolated from this fraction by immunomagnetic selection using a mouse anti-CD31 antibody , followed by an anti-mouse IgG Dynabead, 30 min incubation with each and magnetic separation was performed on MACS columns.
  • Isolated LSECs were…
  • Human liver endothelial cells were isolated from liver tissue from patients undergoing surgical liver resection.
  • The tissue was collected with informed consent and under local ethics committee approval of the University of Bern.
  • Approximately 30 g of liver was cut into 2 mm3 pieces and digested in 50 ml PBS containing 10 mg/ml collagenase 1A for 90 min at 37 °C.
  • The resultant solution was then filtered through a sterile 0.5 mm steel mesh, the supernatant centrifuged at 300 × g for 8 min, and the pellet then resuspended on 10 ml PBS.
  • This digest was then layered on a 33%/77% Percoll density gradient, centrifuged for 30 min at 800 × g 4 °C and the non-parenchymal layer removed, pelleted and resuspended in 500 μl of PBS.
  • Endothelial cells were isolated from this fraction by immunomagnetic selection using a mouse anti-CD31 antibody , followed by an anti-mouse IgG Dynabead, 30 min incubation with each and magnetic separation was performed on MACS columns.
  • Isolated LSECs were cultured on collagen-coated plates.
  • Human LSECs were cultured in SFM endothelial basal media with 10% human serum, 0.1% gentamycin, 5 ng/ml hepatocyte growth factor and 10 ng/ml vascular endothelial growth factor (VEGF, ).

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