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.

ACTIVITY TEST

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.

 

Image


SDS-PAGE Image

 

Figure. Western Blot; Sample: Recombinant CYP1A2, Human.

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