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5 mL iCapTag protein purification column (8 x 100 mm)

02000502R
$1,175.00
In stock
1
Product Details

Description

The i CapTag TM resin is designed to capture recombinant proteins tagged with the i CapTag TM , a self-cleaving affinity tag for the rapid and simple purification of tagless proteins.

Prepacked i CapTag TM columns are designed for simple purification of tagless proteins via a self-cleaving affinity tag.
  • The i CapTag TM resin binds with high specificity to the i CapTag TM , a 36-amino acid sequence appended to the N-terminus of the target protein.
  • Strong and selective binding at pH 8.5 allows purification of the target protein via a conventional affinity purification approach.
  • Once purified, the i CapTag TM is induced to release the target protein from the bound tag via a change in buffer pH from 8.5 to 6.2.
  • Controllable cleavage to 90% released target typically takes place in less than 5 hours at room temperature (depending on the N-terminal amino acids of the target protein). Prescribed modifications of the first two amino acids of the target can be used to optimize cleaving. In some cases, cleaving can take place in minutes.
  • Once the released target protein is recovered in the column effluent, the i CapTag TM resin can be stripped at reused by a low pH wash.
  • The i CapTag TM resin has been used successfully for proteins expressed in E. coli , CHO and HEK cell lines, as well as cell-free systems and non-standard host cells.

Product Specifications

Table 1. Characteristics of the i CapTag TM affinity chromatography.

Parameters

Requirements*

Packing size 1 x 5 mL
Bed dimensions 8 x 100 mm
Bed volume 5 mL
Flow rate** Recommended flow rate is 1.25 mL/min (see manual for more details). Maximum flow rate is 2 mL/min.

Ligand Density

15 ± 2 g/L

Cleavability &

95% or higher cleavage in 5 hours at room temperature (based on GFP test target protein)

Base Bead

Hardened agarose (6% cross-linked agarose)

Particle Size, d 50v

~90 μm

Maximum Back Pressure

40 psi (2.8 bar; 0.28 MPa)

Typical Working Temperature #

4 ℃ to 36 ℃

Chemical Stability

Resin is initially stripped at pH ≤ 2.0 followed by buffers with the pH between 6.2 and 8.5. Resin is neutral to salts, sugars and other buffer additives, including protease inhibitors and stays active when caustic buffers are used.

Resin Packing

50% slurry in storage solvent in 18% ± 2% ethanol

Storage

18% ± 2% ethanol, 4 ℃ to 8 ℃

Abbreviations & Notes

* - Specific parameters of the resin are included in the Certificate of Analysis based on the lot number of the resin.

** - The pressure varies based on column tubing and chromatography medium.

& - Note that the first two amino acids of protein of interest have a significant impact on the cleavage rate, where aromatic residues tend to accelerate cleaving while smaller residues slow or stop cleaving and proline abolishes cleaving (see additional instructions). For proteins with unacceptably slow cleaving, amino acids can be added or modified at the N-terminus to provide more optimal cleaving.

# - Note that the temperature effects cleaving. Typically, lower temperatures slow down cleaving rate and higher speed up cleavage.

d 50v - Median particle size of the cumulative volume distribution.


Safety Information

Ethanol, solution: 18% ± 2%

Hazard class: 3

Identification number: UN1170

Packing Groups: PG 3

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Manual

Download product manual for rapid implementation



Plasmids

Download examples of DNA plasmids containing target proteins

MSDS

Download & Print MSDS

FPLC Columns

MSDS

Download & Print MSDS

Bulk Resin

Certificate

Download the Certificate of Analysis


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