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Human Epidermal Growth Factor Recombinant

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$70.00$220.00

SKU: RKP01133 Tags: , ,

Description

Accession
P01133
Source
Optimized DNA sequence encoding Human EGF mature chain was expressed in Escherichia Coli.
Molecular weight
Recombinant Epidermal Growth Factor is a monomer protein consisting of 54 amino acid residue subunits, migrates as an approximately 6 kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>98%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED50 was determined by a cell proliferation assay using balb/c 3T3 cells is ≤ 0.5 ng/ml, corresponding to a specific activity of ≥2 x 10^7 units/mg.
Protein Sequence
NSDSECPLSH DGYCLHDGVC MYIEALDKYA CNCVVGYIGE RCQYRDLKWW ELR
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
RecombinantEpidermal Growth Factor was lyophilized from a 0.2μm filtered concentrated (1mg/ml) solution in PBS, pH 7.2.
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
Molecular function

Methods

Rodent neurosphere culture, neurosphere differentiation, and grafting

  • Striatum from C57BL/6 mice, transgenic for the GFP gene [strain C57BL/6-TgN(ACTbEGFP)1Osb] (Okabe et al., in vitro differentiation of striatal NSCs could be induced using a simple four-step method.
  • Briefly, after formation of neurospheres (step 2), selection and expansion of ESCs was carried out in medium'>NB27 medium (step 3).
  • Then, neurospheres were dissociated and plated (2 × 105 cells/well) (step 4) into tissue culture dishes pre-coated with polylysine (PLL) and containing NB27 medium without serum or growth factors.
  • Immature NSCs cells were allowed to differentiate for five days in a humidified atmosphere containing 95% air and 5% CO2 at 37°C.
  • After five days, GFP cells were collected by mild trypsinization, suspended in Hanks Balanced Salt Solution (HBSS, Sigma) and the cell suspension adjusted to obtain an appropriate concentration of cells for injection, minutes before transplantation.
  • NSCs obtained either by dissociation by mild trypsinization or neural-differentiated cells obtained…

Neural in vitro differentiation

  • Neural differentiation of LLC6P and LLC9P cells was performed as previously described g for 5 minutes at 4°C and plated on polyornithine/laminin-coated cell culture dishes.
  • Passage number of LLC6P hpESCs at differentiation induction was 30–45; line LLC9P cells were used at passage numbers 48–60.
  • Terminal differentiation of hpNSCs was performed in differentiation media containing DMEM/F12 (N2 supplement; 1∶50) and Neurobasal (B27 supplement; 1∶50) mixed at 1∶1 ratio.
  • cAMP (300 ng/mL) was added to the media for 28 days.
  • For induction of dopaminergic differentiation

Cell culture

  • The human retinoblastoma cell line, WERI-Rb1, obtained from the American Type Culture Collection , was maintained in RPMI-1640 Medium with 10% fetal bovine serum (FBS, , ) [2 and observed under inverted microscopy every other day.

In Vitro Differentiation

  • To examine the ability of hBSCs to spontaneously differentiate, breastmilk cells were initially grown as spheroids (see above).
  • By day 4–7, some cells had attached.
  • The remaining spheroids were transferred into new wells where adherent cells appeared in 1–2 days.
  • Both the initial and subsequent attached cells were cultured for another 2–3 weeks, with media changes every 3–5 days.
  • For directed differentiation, primary and first- to third-passage breastmilk cell cultures were incubated in differentiation media at 37°C and 5% CO2 for 3–4 weeks.
  • For mammary differentiation, cells were incubated in Roswell Park medium'>Memorial medium'>Institute medium (RPMI) 1640 with l-glutamine supplemented with 20% FBS, 4 μg/ml insulin , 20 ng/ml epidermal growth factor (EGF) , 0.5 μg/ml hydrocortisone , 5% antibiotic-antimycotic, and 2 μL/ml fungizone.
  • For osteoblastic and adipogenic differentiation, cells were incubated in NH OsteoDiff or NH Adipodiff medium, respectively .
  • For chondrocyte differentiation,…

Cell Culture

  • H1 (WA01) cells from WiCell were expanded in ES cell growth media and differentiated according to our laboratory’s established protocols EBs) were formed by suspending undifferentiated ES cell colonies in ES growth media on ultra low adhesion culture dishes.
  • The neural differentiation was commenced by growing EBs in medium'>N2B27 medium NPs).
  • NPs were then passaged with 0.05% trypsin/ETA and cultured for 3 weeks in N2B27 media with 20 ng/ml of PGF-AA and 20 ng/ml EGF until transplantation.
  • A human fibroblast line (HFF1, from ATCC) from neonate foreskins were cultured in 10% fetal bovine serum as previously described

Cell culture

  • GL261 glioma cells were cultured in DMEM-F12 Glutamaxâ„¢-1 , B-27 supplement 1X , penicillin/streptomycin 1X, human recombinant epidermal growth factor (EGF; 20 ng/ml), and human recombinant fibroblast growth factor-2 (FGF-2; 20 ng/ml).
  • The spleens and draining lymph nodes of immunized and control mice were surgically removed and mechanically processed in medium'>RPMI medium'>1640 basal medium (LONZA) containing 5% fetal bovine serum, 20 mM HEPES, 2% L-glutamine and penicillin/streptomycin.
  • Erythrocytes were lysed with ice-cold ACK buffer, and lymphocytes were resuspended in RPMI 1640 supplemented with 10% fetal bovine serum, 2% penicillin/streptomycin, 2% L-glutamine, 50 μM β-mercaptoethanol, 100 mM sodium pyruvate and 100X nonessential amino acids.
  • We also added 10 U/ml human recombinant IL-2 to the medium for all in vitro experiments.

Synthesis, purification, spectrum, electrophoresis properties and signaling of EGF-NIR.

  • Insert-12% SDS-PAGE analysis of 10 µg of EGF-NIR scanned with Odyssey and unmodified EGF stained with coomassie blue.

Expression of key growth factors in human endometrium.

  • Brown signals for growth factors (EGF, IGF-1, GM-CSF, BDNF, CSF-1, artemin, and GDNF) were found following staining with specific antibodies.
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