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ACTIVITY TEST
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 > 95% Isoelectric Point 7.7 Applications Cell culture; Activity Assays.
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Anti-Müllerian hormone (AMH), also named Müllerian inhibiting substance (MIS) belongs to a tissue-specific TGF-beta superfamily growth factor. It can be expressed by male sertoli cells and postnatal testis, and ovarian granulosa cells of females postpartum. AMH expression is critical to sex differentiation at a specific time during fetal development, it appears to be tightly regulated by SF1, GATA factors, DAX1 and FSH. AMH signals through a characteristic receptor consisting of a type I and a type II receptor serine/threonine kinase. Especially the type II receptor is unique and specific receptor for AMH. Besides, Mothers Against Decapentaplegic Homolog 9 (Smad9) has been identified as an interactor of AMH, thus a binding ELISA assay was conducted to detect the interaction of recombinant human AMH and recombinant human (Smad9) Briefly, AMH were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to Smad9-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-AMH pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 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 of AMH and Smad9 was shown in Figure 1, and this effect was in a dose dependent manner.
USAGE
Reconstitute in ddH2O 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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Figure. SDS-PAGE
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Figure. Western Blot; Sample: Recombinant AMH, Human.
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