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Recombinant human HGF (NK1) protein (Qk013)
Recombinant human HGF protein is a potent, high purity NK1 isoform of human hepatocyte growth factor (HGF) that promotes efficient differentiation of human iPSCs to hepatocyte-like cells at just 10 ng/ml with highly homogeneous expression of the hepatic marker, HNF4α. The highly scalable animal-free manufacture and enhanced bioactivity make this suitable for chemically-defined media and reproducible scale-up.
20kDa naturally occurring isoform of HGF, animal-free and carrier-protein free.
Orders are typically shipped same or next day (except Friday).
Easy world-wide ordering, direct or through our distributors.
1mg may be despatched as 2 x 500µg
Buy online with secure credit card or purchase order or email orders@qkine.com
Bulk and stock reservation available
Summary
High purity human HGF (NK1) protein (Uniprot: P14210)
>97%, by SDS-PAGE quantitative densitometry
20 kDa
Expressed in E. coli
Animal-free (AOF) and carrier protein-free.
Manufactured in our Cambridge, UK laboratories
Lyophilized from acetonitrile, TFA
Resuspend in 10mM HCl at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C
Featured applications
Differentiation of human iPSCs to hepatocyte-like cells at just 10 ng/ml
1. Organ, Shawna Leslie, and Ming-Sound Tsao. ‘An Overview of the C-MET Signaling Pathway’. Therapeutic Advances in Medical Oncology, vol. 3, no. 1 Suppl, Nov. 2011, pp. S7–19. https://doi.org/10.1177/1758834011422556.
2. Oliveira, Alexandre G., et al. ‘The Role of Hepatocyte Growth Factor (HGF) in Insulin Resistance and Diabetes’. Frontiers in Endocrinology, vol. 0, 2018. Frontiers, https://doi.org/10.3389/fendo.2018.00503.
3. Jakubczak, John L., et al. ‘NK1, a Natural Splice Variant of Hepatocyte Growth Factor/Scatter Factor, Is a Partial Agonist In Vivo’. Molecular and Cellular Biology, vol. 18, no. 3, Mar. 1998, pp. 1275–83. doi: 10.1128/mcb.18.3.1275
Human
Species similarity:
Porcine – 95%
Bovine – 93%
Rat – 90%
Mouse – 89%
Frequently used together
Bioactivity
HGF (NK1) (Qk013) has equivalent bioactivity to a mammalian cell-expressed full-length HGF from another commercial supplier. Comparative bioactivity was determined using a quantitative Promega serum response element luciferase reporter assay in transfected HEK293T cells. Cells were treated (in duplicate) with a serial dilution of HGF for 6 hours. Firefly luciferase activity is measured and normalized to control Renilla luciferase activity.
Qk013 HGF (green) has an EC50 of 49.6 pM (0.99 ng/mL) and the mammalian cell-expressed HGF (black) has an EC50 of 68.4 pM (1.37 ng/mL).
Data from Qk013 lot #010.
Purity
HGF NK1 migrates as major band at 20 kDa in non-reducing conditions and 18 kDa upon reduction.
Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+DTT, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250. NB reduced samples were not boiled as the protein is sensitive to high temperatures, which causes degradation.
Data from Qk013 lot #010
Further quality assays
Mass spectrometry: single species with expected mass
Analytical reversed-phase: single sharp peak
Endotoxin: <0.005 EU/μg protein (below level of detection)
Recovery from stock vial: >95%
We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine. All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.
Protein background
Hepatocyte growth factor (HGF) regulates cell growth, cell motility, and morphogenesis and plays a central role in angiogenesis, tumorogenesis, and tissue regeneration. HGF binds and activates the receptor tyrosine kinase, c-Met; through this receptor HGF activates PI3K/AKT, FAK, JNK, and ERK1/2 signaling.1
HGF is secreted as a single inactive polypeptide and is cleaved by serine proteases into a 62-kDa heavy-chain and 32-36-kDa light-chain. A disulfide bond between the heavy and light chains produces the active, heterodimeric molecule.2 Alternative splicing of HGF produces multiple transcript variants encoding different isoforms. The NK1 isoform is the smallest natuarlly occuring splice variant, it is comprised of the HGF n-terminus and first kringle domain.3
Customer & collaborator data
High purity, animal and carrier protein-free HGF protein for chemically-defined media and more reproducible scale-up
Recombinant human HGF (NK1) protein promotes efficient differentiation of human iPSCs to hepatocyte-like cells
HGF (NK1) supports human iPSC differentiation to hepatocyte-like cells. Defined media was supplemented with Qk013 HGF (NK1) during day 8-18 of the hepatocyte differentiation protocol (adapted from Heck W, et al. 2015). Cells attain characteristic hexagonal epithelial morphology. Expression of hepatic marker, HNF4α, and polarisation marker, E-Cadherin (ECAD), was detected using immunofluorescence.
Highly homogeneous expression of HNF4α in hepatocyte-like cells differentiated from iPSCs using Qk013 HGF (NK1)
Growth in media supplemented with Qk013 HGF (NK1) leads to highly homogeneous expression of hepatic marker, HNF4α. iPSCs were treated with hepatocyte differentiation media supplemented with Qk013 HGF (NK1) or full-length, mammalian cell-derived HGF from another commercial source.
All experiments have been conducted by the specialist stem cell biotechnology company, Stemnovate Limited, in Cambridge, UK.
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
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