Chromatography Matrix

Sepharose CL-2B

Cross-Linked Agarose Gel Filtration Matrix for Affinity Ligand Coupling

Sepharose CL-2B is a well-proven cross-linked agarose gel filtration base matrix frequently used for coupling affinity ligands. The matrix is not pre-activated, allowing the user to perform all steps in coupling.

Cat. No.: Se-500C    |    Pack Size: 1 L
2% Cross-Linked Agarose
60–200 μm Particle Size
pH 3–13 Working Range
1 L Pack Size

Product Overview

Sepharose CL-2B is a well-proven cross-linked agarose gel filtration base matrix and is frequently used for coupling affinity ligands to the matrix. The matrix is not pre-activated and the user performs all steps in coupling.

Sepharose is a bead-formed agarose-based gel filtration matrix. Sepharose is available with 3 different agarose contents; 2%, 4%, and 6%, designated Sepharose 2B, Sepharose 4B and Sepharose 6B respectively.

Both Sepharose and Sepharose CL have broad fractionation ranges which makes them suitable for characterizing or cleaning-up samples containing components of diverse molecular weight.

Key Features

02

2% Cross-Linked Agarose

Bead-formed agarose gel filtration matrix designed for chromatography applications.

AF

Affinity Coupling Support

Proven base matrix for coupling affinity ligands.

ST

Enhanced Stability

Higher degree of crosslinking improves matrix stability.

FR

Broad Fractionation Range

Suitable for characterizing and cleaning-up samples containing components of diverse molecular weight.

PH

Broad pH Range

Working pH range from 3–13 with CIP stability from pH 2–14.

AC

Autoclavable

Autoclavable for 20 min at 120°C in pH 7 phosphate buffer.

Technical Specifications

Parameter Specification
Product Sepharose CL-2B
Cat. No. Se-500C
Matrix 2% cross-linked agarose
Agarose Content 2%
Particle Size 60 μm–200 μm
Recommended Flow Rate <15 cm/h
Recommended Column Height 10 cm
Maximum Operating Pressure 60–120 cm/h
Pressure Drop 15 cm H₂O / bed height
Bed Height 10 cm
Column Diameter 5 cm i.d.
Working pH Range 3–13
CIP Stability 2–14
Autoclavable Autoclavable
Autoclaving Conditions 20 min at 120°C in pH 7 phosphate buffer
Storage 4–30°C, 20% Ethanol
Pack Size 1 L

Applications

Gel Filtration & Size-Based Separation

Suitable for separation and clean-up of biological samples according to molecular size.

Affinity Chromatography

Used as a base matrix for coupling affinity ligands. The matrix is not pre-activated, allowing users to perform the desired coupling chemistry.

Protein Purification

Suitable as a matrix component in chromatography workflows involving proteins and other biological macromolecules.

Sample Characterization

The broad fractionation range can be useful when analyzing samples containing components across different molecular-weight ranges.

Column Equilibration

Before applying the sample, equilibrate the column with at least two column volumes of the eluent to be used in the separation, or until the baseline is stable.

Longer equilibration might be needed with detergent solutions. Equilibration is not needed between runs with the same eluent.

Recommended practice: Allow the baseline to stabilize before sample application to establish consistent chromatographic conditions.

Regeneration

1

Wash

Wash the column with 2–3 column volumes of buffer.

2

Change Conditions

If changing chromatographic conditions, use the new buffer after the washing step.

3

Re-equilibrate

Re-equilibrate the column in the new buffer before the next run.

Sanitization & Sterilization

For sanitization, wash the column with:

0.5 M NaOH

Sepharose CL-2B is autoclavable under the specified conditions.

Autoclaving

20 minutes at 120°C in phosphate buffer, pH 7.

CIP Stability

Stable over the specified CIP range of pH 2–14.

Storage

Temperature

Store at 4–30°C.

Storage Solution

20% Ethanol.

Frequently Asked Questions

What is Sepharose CL-2B?
Sepharose CL-2B is a 2% cross-linked agarose gel filtration matrix used for chromatography and as a base matrix for coupling affinity ligands.
Is Sepharose CL-2B pre-activated?
No. Sepharose CL-2B is supplied as a non-activated matrix. The user performs the required ligand-coupling steps.
What is the particle size?
The specified particle-size range is 60–200 μm.
What is the recommended flow rate?
The recommended flow rate is less than 15 cm/h.
What is the working pH range?
Sepharose CL-2B has a specified working pH range of 3–13.
Can Sepharose CL-2B be autoclaved?
Yes. The matrix is autoclavable for 20 minutes at 120°C in phosphate buffer at pH 7.
How should Sepharose CL-2B be stored?
Store at 4–30°C in 20% ethanol.
How should the column be equilibrated?
Before sample application, equilibrate the column with at least two column volumes of the separation eluent, or until the baseline is stable. Longer equilibration may be required with detergent solutions.
How is the column regenerated?
Regeneration is normally performed by washing with 2–3 column volumes of buffer, followed by re-equilibration when changing conditions.

Sepharose CL-2B

A versatile cross-linked agarose matrix for gel filtration, chromatography, sample clean-up, and affinity ligand coupling workflows.

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Technical Support & Enquiries

United Kingdom Office

Address:
Unicorn House,
Station Close,
Potters Bar EN6 1TL,
United Kingdom

Phone: +44 20 3393 8531

Email: [email protected]