Bio-rad Profinity eXact™ Purification and Tag Cleavage Con Uživatelský manuál stáhnout pdf (Strana 5)

Table 3. Buffer and chemical compatibilities for

Profinity eXact cartridges.

Reagent

Group Description Stability

Triggering Ions F

-

, Cl

-

, N

3

-

, NO

2

-

, CO

2

-

Do not use in lysis and

wash buffers.

See Table 5 for use as

trigger in elution buffer.

Salts Sodium Acetate £3M

NaCl or KCl Do not use in lysis and

wash buffers.

Buffers Tris-HCl Substitute Tris-acetate

or Tris-phosphate.

Acids HCl Do not use. Substitute

acetic or phosphoric

acid.

Detergents Non-ionic £5% (w/v)

Zwitterionic £5% (w/v)

Ionic Do not use.

Protease PMSF, CalBioChem cocktail,

inhibitors Roche Protease Inhibitor 1X

cOmplete mini tablet

BD Biosciences cocktail 2X

Lysis Solutions Lysis & Extractrion Reagent 1X

(Bio-Rad), B-PER phosphate

(Pierce), BugBuster (EMD),

FastBreak Cell Lysis (Promega)

ReadyPreps Lysis (Epicentre), Do not use.

CelLytic Express (Sigma)

5

Table 2. Profinity eXact resin specifications.

Functional ligand Mutant subtilisin

Base bead Superflow 6% agarose

Particle size range 60–160 µm

Recommended linear flow rate <1,000 cm/hr at 25°C

Maximum operating pressure 45 psi

Chemical compatibility See Table 3

Storage 4°C

Shelf life in storage buffer >1 year at 4°C

Operational temperature 4–40°C

4

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