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IDA Agarose for Phosphoproteins & Phosphopeptides

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Description

FastSep high performance resins and chromatography cartridges

Numerous ligand-metal ion combinations 
High affinity & high capacity 
Minimal non-specific binding & excellent purity 
Long spacer arms & minimal steric hindrance 
Low metal ion leaching & excellent stability 
Maintain high performance after multiple repeated use 

 

Application

For phosphoprotein & phosphopeptide purification 
Simply replace your existing NTA resin or cartridge, no optimization or protocol changes necessary 
Purification under native and denaturing conditions 
Suitable for small proteins, large protein complexes, and proteins with low expression rates 


Purification of Phosphoproteins & Phosphopeptides

Immobilized Al3+, Ga3+, and Fe3+ are the most widely used ions for selective enrichment of phosphopeptides from complex proteolytic digest mixtures containing both phosphorylated and nonphosphorylated components. The most commonly used ligands are NTA and IDA. Depending on identity of proteins, impurity profile and purification conditions, different ligand-metal ion combinations can achieve different purification results. 

 

How to choose different ligands and metal ions in IMAC?

NTA and IDA are the two most widely used ligands in IMAC. NTA is a tetravalent ligand with coordination number 4 and therefore stronger coordination of metal ions, while IDA is a trivalent ligand with coordination number 3. Generally, NTA gives higher binding specificity and lower metal ion leaching, while IDA shows more non-specific binding and increased metal ion leaching. IDA can have higher metal ion loading density and usually requires much lower imidazole concentration in eluent. 


As many factors can affect the choice of optimal ligand-metal ion combination, the best approach is to try different combinations and let your proteins decide. FastSep(TM) high performance resins and cartridges offer numerous ligand-metal ion combinations to meet various purification needs. 


FastSep(TM) high performance resins and chromatography cartridges are based on highly cross-linked agarose 6% beads which have structural characteristics that result in superb physical strength, excellent flow properties, low backpressure, and an open pore structure. FastSep(TM) high performance resins and chromatography cartridges allow operation at very high flow rates, resulting in increased throughput and rapid column cleaning between chromatography runs.

 

Name

Cat#

Description (every combination is 50% suspension)

Al-IDA Resin

ALIDA-25

25ml aluminum-charged IDA resin (50ml total volume)

Al-IDA Resin

ALIDA-100

100ml aluminum-charged IDA resin (200ml total volume)

Al-IDA Resin

ALIDA-500

500ml aluminum-charged IDA resin (1000ml total volume)

Ga-IDA Resin

GAIDA-25

25ml gallium-charged IDA resin (50ml total volume)

Ga-IDA Resin

GAIDA-100

100ml gallium-charged IDA resin (200ml total volume)

Ga-IDA Resin

GAIDA-500

500ml gallium-charged IDA resin (1000ml total volume)

Fe-IDA Resin

FEIDA-25

25ml iron-charged IDA resin (50ml total volume)

Fe-IDA Resin

FEIDA-100

100ml iron-charged IDA resin (200ml total volume)

Fe-IDA Resin

FEIDA-500

500ml iron-charged IDA resin (1000ml total volume)

XX-IDA Resin

XXIDA-OEM

Any size any metal ion charged IDA resin

 

Specification 

Chromatography technique

Phosphoprotein & Phosphopeptide purification

Matrix 

Highly cross-linked 6% beaded agarose

Active group

Al3+, Ga3+ or Fe3+ charged Iminodiacetic Acid (IDA)

Active group density

>40 umol/ml drained medium 

Binding capacity

>40 mg/ml drained medium

Spacer 

12 atom stable and uncharged ether hydrophilic linkage

Bead geometry & size

Spherical 50 to 150 um

Bead mean diameter d50v

90 um 

Linear flow velocity

<75 cm/h at 25 °C, HR 16/10 column, 5 cm bed height 

Recommended linear flow rate

<30 cm/h

Pressure/flow specification

Base matrix 100-200 cm/h, pressure drop cm H2O/bed height = 15, bed height 10 cm, 5 cm i.d. 

1ml Cartridge bed dimensions

7×25 mm

1ml Cartridge bed height

25 mm 

1ml Cartridge bed volume

1 ml 

1ml Cartridge column I.D

7 mm

5ml Cartridge bed dimensions

16×25 mm

5ml Cartridge bed height 

25 mm

5ml Cartridge bed volume 

5 ml 

5ml Cartridge column I.D

16 mm

Maximum cartridge flow rate

5 ml/min (1 ml cartridge) or 20 ml/min (5ml cartridge) 

Recommended cartridge flow rate  

1 ml/min (1 ml cartridge) or 5 ml/min (5 ml cartridge)

Maximum pressure during operation

5 bar [0.5 MPa] (70 psi)

pH Stability working range

3 to 13 

pH Stability cleaning in place (cip)

2 to 14

Chemical stability

Stable to commonly used aqueous solutions. Can be used with non-ionic detergents, denaturing solvents, e.g. 8 M urea and 6 M guanidine hydrochloride, Stable in organic solvents, e.g. 50% methylformamide and 50% dioxane.

Storage

2 to 8 °C

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