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	> 90%
Isoelectric Point	9.4
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Figure 1. The binding activity of CYP3A4 with HSPA8.
Cytochrome P450 3A4 (CYP3A4) is an important enzyme in the body, mainly found in the liver and in the intestine. It oxidizes small foreign organic molecules (xenobiotics), such as toxins or drugs, so that they can be removed from the body. CYP3A4 is a member of the cytochrome P450 family of oxidizing enzymes and the CYP3A4 is the most common and the most versatile one in drug metabolism. Besides, Heat Shock 70kDa Protein 8 (HSPA8) has been identified as an interactor of CYP3A4, thus a binding ELISA assay was conducted to detect the interaction of recombinant human CYP3A4 and recombinant human HSPA8. Briefly, CYP3A4 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to HSPA8-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CYP3A4 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 CYP3A4 and HSPA8 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


Figure. SDS-PAGE

 

Figure. Western Blot

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