Gallus Active Transforming Growth Factor Beta 2 (TGFb2)

Instruction Manual!

Product name:	Gallus Active Transforming Growth Factor Beta 2 (TGFb2)
Source:	Prokaryotic expression
Purity:	>92%
Buffer Formulation:
Applications:	Cell culture; Activity Assays.
Storage:	Avoid repeated freeze/thaw cycles. Store at 2-8oC for one month. Aliquot and store at -80oC for 12 months.
UOM:

[ PROPERTIES ]

Source: Prokaryotic expression.

Host: E. coli

Residues: Leu279~Ser412

Tags: N-terminal His-tag

Purity: >92%

Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05%

sarcosyl and 5% trehalose.

Applications: Cell culture; Activity Assays.

(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 8.9

Predicted Molecular Mass: 19.1kDa

Accurate Molecular Mass: 19kDa as determined by SDS-PAGE reducing

conditions

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0

mg/mL. Do not vortex.

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles.

Store at 2-8oC for one month.

Aliquot and store at -80oC for 12 months.

Stability Test: 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 37oC 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.

[ SEQUENCE ]

[ ACTIVITY ]

Transforming growth factor beta (TGF-β) is a multifunctional cytokine belonging to

the transforming growth factor superfamily. The TGF- β superfamily includes

endogenous growth inhibiting proteins; an increase in expression of TGF- β often

correlates with the malignancy of many cancers and a defect in the cellular growth

inhibition response to TGF- β . Its immunosuppressive functions then come to

dominate, contributing to oncogenesis. To test the effect of TGF-β on inhibit HGF-

dependent proliferation, HepG2 cells were seeded into triplicate wells of 96-well

plates at a density of 2,000 cells/well and allowed to attach, replaced with serum-

free overnight, then the medium was replaced with 2% serum standard DMEM

including 1ng/mL HGF prior to the addition of various concentrations of

recombinant chicken TGF- β . After incubated for 96h, 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

the absorbance at 450nm was measured using a microplate reader after incubating

the plate for 1-4 hours at 37℃. The inhibitory effect of TGF-β on HGF-dependent

proliferation of HepG2 cells observed by inverted microscope was shown in Figure1.

Cell viability was assessed by CCK-8 assay after incubation with recombinant TGF-

β for 96h. The result was shown in Figure 2. It was obvious that TGF-β significantly

decreased cell viability of HepG2 cells.

Figure 1. The inhibitory effect of TGF-β on cell proliferation of HepG2 cells.

(A) HepG2 cells cultured in DMEM, stimulated with 1µg/mL TGF-β for96h;

(B) Unstimulated HepG2 cells cultured in DMEM for 96h

Figure 2. TGF-β inhibit cell proliferation of HepG2 cells.

[ IDENTIFICATION ]

Figure 3. SDS-PAGE

Sample: Active recombinant TGFb2, Gallus

Figure 4. Western Blot

Sample: Recombinant TGFb2, Gallus;

Antibody: Rabbit Anti-Gallus TGFb2 Ab