Bio-Rad Bio-Gel P Polyacrylamide Gel User Manual
Bio-Gel®P
Polyacrylamide Gel
Instruction Manual
Table of Contents
Section 1 Introduction.............................................................1
Section 2 Technical Description .............................................3
Section 3 Instructions for Use ................................................6
3.1 Column Selection................................................................6
3.2 Eluant Selection...................................................................6
3.3 Preparation of the Gel..........................................................8
Section 4 Sample....................................................................12
Section 5 Void Volume Determination and Calibration....14
Section 6 Sanitation and Sterilization .................................15
Section 7 Storage...................................................................15
Section 8 Flow Rate Determination.....................................16
Section 9 Ordering Information ..........................................18
Section 1
Introduction
Bio-Gel P gels are porous polyacrylamide beads prepared by
copolymerization of acrylamide and N,N'-methylene-bis-acrylamide.
The gels are extremely hydrophilic and essentially free of charge,
and provide efficient, gentle gel filtration of sensitive compounds.
Their synthetic composition and freedom from soluble impurities
preclude eluate contamination. High resolution is assured by consistent narrow distribution of bead diameters and excellent molecular
weight discrimination.
Bio-Gel P gel is compatible with dilute organic acids, 8 M urea,
6 M guanidine-HCl, chaotropic agents, reducing agents such as dithiothreitol and mercaptoethanol, and detergents such as SDS, CHAPS,
and Triton
tilled water however, buffers of > 50 mM ionic strength are recommended for most protein separations.
Bio-Gel P gel. Alcohol up to 20% will not substantially alter the
®
X-100. Bio-Gel P gel may be used effectively with dis-
Miscible organic solvents may be added to the eluants used with
1
exclusion properties of the gel, and will in some cases enhance
separation of complex mixtures of poorly water soluble small
molecules such as nucleotides, peptides, and tannins. Formamide
may be used at full strength, because Bio-Gel P gel is completely
swelled by this solvent.
Bio-Gel P gel is autoclavable at pH 5.5-6.0 in buffers such as
50 mM HEPES, MES, or citrate at 120 °C for 15-30 minutes. At
room temperature, the recommended operating pH range is 2-10.
Bio-Gel P gel is susceptible to hydrolysis of amide groups at higher
or lower pH. Flow rate and resolution increase with temperature over
the range of 4-80 °C.
Section 2
Technical Description
Table 1. Bio-Gel P Gel Product Description
Matrix Bio-Gel polyacrylamide gel
Particle size
Medium 90-180 µm
Fine 45-90 µm
Extra fine < 45 µm
Shipping medium Shipped dry
Resistance
pH 2-10
Pressure 15 psi
Organic solvents < 20%
Working temperature range 4-80 °C
Temperature limits Autoclavable, at pH 5.5-6.5, at
120 °C for 30 min
Storage Dry, at r oom temperature; in dis-
tilled water or aqueous buffers
at 4 °C with 0.02% sodium azide
2
3
Table 2. Properties of Bio-Gel P-Gels
Particle Size Hydrated Bed Range/Nominal
Gel Beads (µM) of Dry Gel Rates (cm/hr)* (Daltons)**, †
Bio-Gel P-2 Gel, Fine 45-90 3 5.0-10 100-1,800
Bio-Gel P-2 Gel, Extra Fine < 45 <10 100-1,800
Bio-Gel P-4 Gel, Medium 90-180 4 15-20 800-4,000
Bio-Gel P-4 Gel, Fine 45-90 10.0-15 800-4,000
Bio-Gel P-4 Gel, Extra Fine < 45 <10 800-4,000
Bio-Gel P-6 Gel, Medium 90-180 6.5 15-20 1,000-6,000
Bio-Gel P-6 Gel, Fine 45-90 10.0-15 1,000-6,000
Bio-Gel P-6 Gel, Extra Fine < 45 <10 1,000-6,000
Bio-Gel P-6DG Gel 90-180 6.5 15-20 1,000-6,000
Bio-Gel P-10 Gel, Medium 90-180 7.5 15-20 1,500-20,000
Bio-Gel P-10 Gel, Fine 45-90 10.0-15 1,500-20,000
Bio-Gel P-30 Gel, Medium 90-180 9 7.0-13 2,500-40,000
Bio-Gel P-30 Gel, Fine 45-90 6.0-11 2,500-40,000
Bio-Gel P-60 Gel, Medium 90-180 11 4.0-6 3,000-60,000
Bio-Gel P-60 Gel, Fine 45-90 3.0-5 3,000-60,000
Bio-Gel P-100 Gel, Medium 90-180 12 4.0-6 5,000-100,000
Bio-Gel P-100 Gel, Fine 45-90 3.0-5 5,000-100,000
Range, Hydrated Volume, ml/g Typical Flow Exclusion Limit
Typical Fractionation
Typical
* Flow rates determined in a 1.5 x 70 cm
column, using a hydrostatic pressure
head:bed of 1:1.
** Fractionation ranges above 40,000 dal-
tons are for globular molecules.
† For quality control purposes, the exclu-
sion limits are determined by calculating
the Kd, or distribution coefficient. The
distribution coefficient is a measure of
the residence time of a molecule in the
pores of the gel, and is expressed as:
(V
- Vo)/(Vt- Vo), where Veis the elution
e
volume of the individual proteins, V
the void volume and V
able volume measured by a small
molecule such as vitamin B
is the total avail-
t
is
o
.
12
4
5
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