///Mouse Epidermal Growth Factor Recombinant

Mouse Epidermal Growth Factor Recombinant



SKU: RKP01132 Tags: ,


Optimized DNA sequence encodingMouse EGF mature chain was expressed in Escherichia Coli.
Molecular weight
RecombinantMouse EGF, generated by the proteolytic removal of the signal peptide and propeptide, is a monomer protein. Recombinant Mouse EGF consists of 54 amino acidsandmigrates as an approximately6.2 kDa protein under reducing conditions.
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED50 was determined by a cell proliferation assay using balb/c 3T3 cells is ≤ 0.1 ng/ml, corresponding to a specific activity of ≥ 1 x 107 units/mg.
Protein Sequence
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Mouse EGF was lyophilized from a 0.2 μm filtered solution in PBS, pH 7.5.
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
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.
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Molecular function


Cell Culture

  • Mouse NS cells containing a constitutively expressed marker for green fluorescent protein (GFP) were used to investigate the process of neural differentiation.
  • Primary cells were obtained from central nervous system tissue of embryonic mice, and cultured primarily as previously described 2.
  • Subculture occurred at day 7 with mechanical dissociation of neurospheres.
  • The differentiation of mouse NS cells in vitro was induced by culturing dissociated cells in differentiation medium containing MEM/F12 with 1× N-2 supplement, 100 ng/ml bFGF, 10% fetal bovine serum (FBS) , 500 nM all-trans retinoic acid , 50 µM taurine , 10 ng/ml transforming growth factor-β2 (TGF-β2) and 1% penicillin-streptomycin in tissue culture plates pre-coated with poly--lysine .
  • The culture medium was changed every 3 days.
  • Differentiated cells at 5×105 cells/ml were processed for immunobloting assays.
  • Transfections and cell treatments were carried out at 2.5×105 cells/ml cellular density.

Hepatocyte isolation and culture

  • Hepatocytes were isolated from the whole liver of an adult ICR mouse (male, 7–8 weeks old) by the two-step liver perfusion method of45 H2O , 50 pM ZnSO47 H2O

Tissue culture

  • Tumors derived from mice deficient in both p53 and patched were kindly provided by Dr. James A. Waschek at UCLA

Neurosphere culture

  • Neurospheres were generated from cells isolated from the subventricular zone of 6–8 week old C57BL/6 mice and cultured in proliferation media consisting of DMEM/F12 (GibcoBRL/Invitrogen) containing 0.6% glucose, 3 mM NaHCO3, 5 mM HEPES, 2 mM L-glutamine, 0.1 mg/ml apo-transferrin, 25 µg/ml insulin, 60 µM putrescene, 30 mM sodium selenite, 20 nM progesterone and 1% BSA (all from), supplemented with EGF (20 ng/ml) and FGF2 (10 ng/ml), as previously described

Neurosphere cultures

  • Neurosphere (NS) cultures were derived from adult mice (∼ four month-old).
  • Briefly, 4 mice per culture were decapitated, brains removed and the hippocampus and subventricular zone were dissected, minced and dissociated with DMEM containing glutamine, gentamicin and fungizone.
  • After treatment with trypsin-EDTA, hialuronidase and DNAse, myelin was removed by using DPBS .
  • Cells were seeded into six-well dishes and cultured in DMEM:F12 (1∶1) containing 20 ng/ml epidermal growth factor , 20 ng/ml fibroblast growth factor (FGF, ) and B27 medium .
  • After ∼10 days in culture, some NS were centrifuged five min at 1000× g, washed with phosphate-buffered-saline and protein was extracted in RIPA buffer.
  • Twenty µg of total protein was loaded on a 10% SDS-PAGE gel and western blot analysis to detect C/EBPβ was performed as previously described C/EBPβ and C/EBPβ to produce neurons, NS from 10-day old cultures were plated for 72…

Soluble amyloid precursor protein alpha (sAPPα) enhances proliferation in an EGF/bFGF-independent manner.

  • (b) A neurosphere formation assay was performed without growth factors (EGF and bFGF), and NPCs were treated with an inactive inhibitor , GM6001 , or GM6001 + recombinant sAPPα (GM+rec sAPPα).

Isolation of intestinal crypts

  • Tissue was removed from PBS solution and washed multiple times with ice-cold PBS washes until the solution remained clear.
  • The specimen was then placed in a Petri dish containing PBS on ice with the mucosal surface facing upward.
  • Using a razor blade, excess mucoid material was scrapped from the epithelial surface.
  • The specimen was then divided into approximately 0.5 cm2 pieces.
  • These pieces were placed into a 2.5 mmol/L EDTA solution in PBS for 30 minutes of incubation with gentle shaking at 4°C.
  • After this incubation period, the fragments were allowed to settle and the supernatant was discarded.
  • 10 ml of cold PBS was added to the sample, and subsequently vortexed for 10 seconds with 1-second bursts.
  • The fragments were allowed to settle, and the supernatant was removed and saved on ice.
  • Again 10 ml of PBS were added and the process was repeated eight times.
  • Samples were spun down at 100 g for 2 minutes.
  • The supernatant…

Cell culture

  • Hepatic stellate cells, Kupffer cells and endothelial cells were isolated from Mx-conditional knockout mice (Mx-cKO) which carry the Mx-promoter attached to cre and are homozygous for the floxed fibronectin gene in vivo prior to cell isolation as described 1 (1-10 ng/ml), murine platelet derived growth factor-BB (PDGF-BB) (100 ng/ml) (Sigma-Adrich) or murine epidermal growth factor (EGF) (100 ng/ml) .
  • The TGF-β signaling inhibitor (SB431542) was used at a final concentration of 10 µM 1, PDGF-BB or EGF lasted for 24 hours or as described in the figure legends, and treatment with the inhibitor lasted one hour, after which TGF-β was added and left for an additional 2 hours before collecting the samples.
  • Fibronectin used for coating was isolated from outdated human plasma as described

Pancreatic Cell Culture

  • Digested pancreatic cells were seeded on a gelatin-coated 6-well plate at 5×104 cells/cm2.
  • Our standard culture medium is a 1∶1 mixture of medium'>Dulbecco's medium'>modified medium'>Eagle's medium and F-12 with 5% fetal bovine serum , Nicotinamide (10 µmol/l), β-mercaptoethenol (50 µmol/l), HEPES (5 mmol/l), and Gentamycin (50 µg/ml).
  • At 24 h after seeding, the medium was changed to the standard medium supplemented with epidermal growth factor (EGF) (20 ng/ml), glucagon-like peptide (GLP)-1 (10 ng/ml), and γ-insulin (10 µM).
  • At day 12 of culture, cultured cells were used for RT-PCR analysis, histological analysis, and transplantation.
  • For the histological analysis of frozen sections and transplantation, cultured cells were detached from the culture dish by pipetting with 2 mg/ml collagenase type V/PBS, taking care not to disrupt the 3D structure of islets and ducts formed in vitro.

Characteristics of fetal radial glia-like cell clones.

  • Cell viability was determined by MTT-assay in cultures maintained with EGF (20 ng/ml), with the EGF receptor antagonist AG 1478 (10−7 M) or with both (e).
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