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Active Mothers Against Decapentaplegic Homolog 2 (Smad2)

JV18; JV18-1; MADH2; MADR2; HMAD-2; HSMAD2; Mad-related protein 2

ACTIVITY TEST

Mothers Against Decapentaplegic Homolog 2 (Smad2) is a pivotal intracellular mediator of the TGF-β signaling pathway that regulates diverse cellular processes including proliferation, differentiation, and apoptosis. As an R-Smad protein, Smad2 transduces signals from activated TGF-β receptors to the nucleus, where it modulates gene expression in complex with Smad4. Smad2 is essential for embryonic development and tissue homeostasis, with its dysfunction linked to cancer and fibrotic diseases. Following TGF-β stimulation, Smad2 binds to and is phosphorylated by the activated Transforming Growth Factor Beta Receptor I (TGFbR1), initiating downstream signaling cascades.To detect the activity of recombinant Smad2, a functional ELISA assay was performed to evaluate the interaction between recombinant human Smad2 and recombinant human TGFbR1.Briefly, biotin-linked Smad2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to TGFbR1-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of Smad2 and TGFbR1 was shown in Figure 1, the EC50 for this effect is 0.77µg/mL.

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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