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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 8.0 Applications Cell culture; Activity Assays.
CYP1A2 (Cytochrome P450 1A2) belongs to the group of proteins which contains heme as a cofactor. CYP1A2 oxidizes a variety of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics. Besides, ASAH1 (Acid ceramidase) has been identified as an interactor of CYP1A2 through affinity capture-MS. Thus a binding ELISA assay was conducted to detect the interaction of recombinant human CYP1A2 and recombinant human ASAH1. Briefly, CYP1A2 were diluted serially in PBS, with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to ASAH1-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-CYP1A2 mAb, 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 CYP1A2 and ASAH1 was shown in Figure 1, and this effect was in a dose dependent manner.
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.
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SDS-PAGE Image
Figure. Western Blot; Sample: Recombinant CYP1A2, Human.
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