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	> 97%
Isoelectric Point	6.6
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Transforming growth factor alpha (TGF-α), a ligand for the epidermal growth factor receptor, which activates a signaling pathway for cell proliferation, differentiation and development. To test the effect of TGF-α on cell proliferation of 3T3 fibroblasts, 3T3 cells were seeded into triplicate wells of 96-well plates at a density of 2, 000 cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM prior to the addition of various concentrations of TGF-α. 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 TGF-α for 72h observed by inverted microscope was shown in Figure 1.

The dose-effect curve of TGF-α was shown in Figure 2. It was obvious that TGF-α significantly promoted cell proliferation of 3T3 cells. The ED50 for this effect is typically 0.6198 to 8.210ng/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


SDS-PAGE Image

 

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

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