Buffer Formulation	20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300.
Traits	Freeze-dried powder
Purity	> 90%
Isoelectric Point	4.8
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Epidermal growth factor (EGF) is a growth factor that stimulates cell growth, proliferation, and differentiation by binding to its receptor EGFR. To test the effect of EGF on cell proliferation of 3T3 fibroblasts, 3T3 cells were seeded into triplicate wells of 96-well plates at a density of 2, 000cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM prior to the addition of various concentrations of EGF. After incubated for 72h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10µL of CCK-8 solution was added to each well of the plate, then measure the absorbance at 450nm using a microplate reader after incubating the plate for 1-4 hours at 37°C .
Cell proliferation of 3T3 cells after incubation with EGF for 72h observed by inverted microscope was shown in Figure 1.

The dose-effect curve of EGF was shown in Figure 2. It was obvious that EGF significantly promoted cell proliferation of 3T3 cells. The ED50 for this effect is typically 0.9848 to 2.958ng/mL.

 

USAGE

 

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

 

STORAGE

 

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

 

STABILITY

 

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

 

Image


Figure. Western Blot; Sample: Recombinant EGF, Human.

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