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Active Tumor Necrosis Factor Beta (TNFb)

LT; LTA; TNF-B; TNFSF1; Tumor Necrosis Factor Ligand Superfamily Member 1; Lymphotoxin Alpha

ACTIVITY TEST

TNF-β, a member of the tumor necrosis factor family, is a potent lymphoid factor that exerts cytotoxic effects on a wide range of tumor cells . The biological effects of TNF-β are very similar to TNF-α,due to the similarity of molecular structure and the receptors.As reported, TNF-α could inhibit the proliferation and induce apoptosis of A549 cells,therefore, A549 cells were seeded into triplicate wells of 96-well plates at a density of 4,000 cells/well with 5% serum standard DMEM including various concentrations of recombinant rat TNF-β. After incubated for 48h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 µL of CCK-8 solution was added to each well of the plate, then the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 2 hours at 37℃. Proliferation of A549 cells after incubation with TNF-β for 48h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with recombinant mouse TNF-β for 48h. The result was shown in Figure 2. It was obvious that TNF-β significantly inhibit cell viability of A549 cells. The ED50 is 2.5μg/ml.

Figure 2. Inhibition of A549 cells proliferation after stimulated with TNF-β

USAGE

Reconstitute in 10mM PBS (pH7.4) 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.

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