Guide to His affinity purification products

 

Scientists at Cube Biotech purify his-tagged proteins almost every day. New ideas how to optimize the procedure lead to novel products with new exciting features. However, it is difficult to keep track on new developments, and to decide which product to choose for a given application. Therefore, we have built two overview tables for Agaroses and MagBeads that -we hope- are helpful.

Please do not hesitate to contact us directly if you are still unsure, or if you are interested in customized or bulk products.

 

 

Table 1: Cube Biotech His Affinity Agarose products.

 

   

His Affinity Agarose

 

PureCube 100
INDIGO

Ni-Agarose

 

 

PureCube 100
Ni-NTA
Agarose

  PureCube
Ni-NTA
Agarose
  PureCube
Ni-NTA
Agarose

XL

 

PureCube
Ni-IDA
Agarose

 
   

Base material

  PureCube 100 Agarose  

PureCube 100 Agarose

 

PureCube Agarose

  PureCube Agarose XL   PureCube Agarose
    Bead diameter   50-150 µm   50-150 µm  

20-50 µm

  300-500 µm   20-50 µm
   

Ligand

  Proprietary   NTA   NTA   NTA   IDA
   

Binding capacity

  up to 80 mg/ml   up to 80 mg/ml   up to 80 mg/ml   >20 mg/ml   50 mg/ml
   

Purity

  High   High   High   High   Medium
   

EDTA tolerance

  20 mM   1 mM   1 mM   1 mM   up to 1 mM
   

DTT tolerance

  20 mM   10 mM   10 mM   10 mM   up to 10 mM
   

pH tolerance

  4-9   2-13   2-13   2-13   2-13
   

Cleaning in place

 

 

GuHCl, Urea, Ethanol

 

NaOH,

GuHCl, Urea, Ethanol

 

NaOH,

GuHCl, Urea, Ethanol

 

NaOH,

GuHCl, Urea, Ethanol

 

NaOH,

GuHCl, Urea, Ethanol

   

Metal stripping

  No   Yes   Yes   Yes   Yes
   

Flow rate (batch/FPLC)

  Fast   Fast   Medium   n.a.   Medium
   

Available metal ions

  Ni   Ni, Co   Ni, Co, Al
Cu, Fe, Zn
  Ni   Ni
   

Special feature

 

NEW

High flow rates, EDTA-stable,

compatible

with eukaryotic growth media

 

NEW

High flow rates, High yields,

High purity

 

Standard material,

available with various

metal ions

 

For special applications

  The economic alternative

 

 

Table 2: Cube Biotech His Affinity MagBead products.

 

   

His Affinity MagBeads

 

PureCube
INDIGO

Ni-MagBeads

  PureCube
Ni-NTA
MagBeads
   

PureCube
Ni-NTA
MagBeads

XL
  PureCube
Ni-IDA
MagBeads

 

   

Base material

 

PureCube

MagBeads

 

PureCube

MagBeads

   

PureCube

MagBeads XL

 

PureCube

MagBeads

    Bead diameter   25 µm   25 µm     70-120 µm   25 µm
   

Ligand

  Proprietary   NTA     NTA   IDA
   

Binding capacity

  up to 80 mg/ml   up to 80 mg/ml     >20 mg/ml   >50 mg/ml
   

Purity

  High   High     High   Medium
   

EDTA tolerance

  20 mM   1 mM     1 mM   up to 1 mM
   

DTT tolerance

  20 mM   10 mM     10 mM   up to 10 mM
   

pH tolerance

  4-9   2-13     2-13   2-13
   

 

Compatible cleaning agents
 

 

GuHCl, Urea,

Ethanol

 

NaOH,

GuHCl, Urea,

Ethanol

   

NaOH,

GuHCl, Urea,

Ethanol

 

NaOH,

GuHCl, Urea,

Ethanol

   

Metal stripping with EDTA

  No   Yes     Yes   Yes
   

Magnetic separation

  Fast   Fast     Medium   Fast
   

Available metal ions

  Ni   Ni, Co, Al
Cu, Fe, Zn
    Ni   Ni
   

Special feature

 

NEW

EDTA-stable, compatible

with eukaryotic growth media.

 

High yield & purity,

Broad ion variety

 

   

For special applications

Larger beads

on request

 

  The economic alternative