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	7.4
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Perforin 1 (PRF1) is a pore forming cytolytic protein found in the granules of cytotoxic T lymphocytes (CTLs) and NK cells. Upon degranulation, perforin binds to the target cell's plasma membrane, and oligomerises in a Ca2 dependent manner to form pores on the target cell. The pore formed allows for the passive diffusion of a family of pro-apoptotic proteases, known as the granzymes, into the target cell. The activity of recombinant PREF1 was measured by lysis of erythrocytes using a hemolysis assay. A general procedure is as fllows: two-fold dilute the recombinant mouse PREF1 with 0.9% NaCl, add 50μl a serial dilution of PREF1, 10μl 0.1M CaCl2 to each well, then add 50μl 0.25% rabbit erythrocyte (RaE) to each well and mixed gently. Add 50μl 0.9% NaCl to reaplace PREF1 in control wells. The plate is incubated for 20 hours at 37 ℃, 5% CO2. The results are shown in Figure 1. It was obvious that the minimal effective concentration of PREF1 is 2.5μg/ml.

 

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 . Gene Sequencing (extract)

Image


Figure. SDS-PAGE

 

Figure. Western Blot

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