Thermo Scientific Pierce Thiophilic Adsorbent is a protein-free immunosorbent resin for gentle preferential binding and purification of immunoglobulins from a variety of sources including mouse, rat, rabbit, goat and human.
Pierce Thiophilic Adsorbent is 6% beaded agarose that has been modified to contain simple sulfone-thioether groups. The result is an affinity resin that has a high binding capacity and a broad specificity for many species of immunoglobulins when incubated in 0.5M potassium sulfate. In effect, thiophilic adsorption is a resin-based variation of ammonium sulfate precipitation for quick immunoglobulin purification from crude serum samples. Unlike antibody fractionation by ammonium sulfate precipitation, purification with thiophilic adsorbent yields concentrated, essentially salt-free, highly purified immunoglobulins at near-neutral pH. Thus, this simple one-step method eliminates the need for additional treatment of the sample for storage or for subsequent conjugation reactions. The gentle binding and elution conditions ensure a high protein recovery with excellent preservation of antibody activity. The resin is available as both a standalone product and as a purification kit containing pre-packed columns and ready-to-use buffers.
Highlights:
- High capacity – gently bind and purify approx. 20mg polyclonal antibody per milliliter of adsorbent resin
- Broad antibody specificity – effective for most immunoglobulin classes and isotypes derived from many different animal species, including chicken IgY
- Simple and fast – quickly fractionate and purify the immunoglobulin component of serum or ascites samples without subsequent dialysis or desalting steps
- Gentle – binding condition is nondenaturing and the elution buffer yields concentrated, salt-free immunoglobulin at near neutral pH; provides excellent recovery of functional antibody
Product Details:
Thiophilic adsorption is based on the ability of some proteins, particularly immunoglobulins, to bind to an immobilized ligand that contains a sulfone group in close proximity to a thioether. Salts that interact with water molecules, such as potassium and ammonium sulfate, promote this binding by driving the protein into close proximity with the ligand.
| Antibody yields and purity levels using Thermo Scientific Pierce Thiophilic Adsorbent. |
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Species
|
Total A280 Bound
|
% Purity by HPLC
|
|
Human
|
4.8
|
70
|
|
Mouse Serum
|
8.6
|
63
|
|
Mouse IgG1
|
11.6
|
92
|
|
Mouse IgG2a
|
9.3
|
88
|
|
Mouse IgG2b
|
9.8
|
97
|
|
Mouse IgG3
|
10.7
|
94
|
|
Rat
|
13.0
|
79
|
|
Bovine
|
17.9
|
90
|
|
Calf
|
11.1
|
89
|
|
Chicken
|
5.2
|
76
|
|
Dog
|
12.2
|
91
|
|
Goat
|
17.3
|
92
|
|
Guinea Pig
|
11.1
|
71
|
|
Horse
|
13.0
|
93
|
|
Pig
|
21.1
|
90
|
|
Rabbit
|
6.7
|
84
|
|
Sheep
|
12.3
|
89
|
References:
- Palmer, D.A., et al. (1994). Flow injection immunoassay for albumin using thiophilic gels. Anal. Biochem., 222, 281-283.
- Porath, J., Maisano, F. and Belew, M. (1985). Thiophilic adsorption – a new method for protein fractionation. FEBS Lett. 185, 306-310.
- Hutchens, T.W. and Porath, J. (1986). Thiophilic adsorption of immunoglobulins – analysis of conditions optimal for selective immobilization and purification. Anal. Biochem. 159, 217-226.
- Belew, M., Juntti, N., Larsson, A. and Porath, J. (1987). A one-step purification method for monoclonal antibodies based on salt-promoted adsorption chromatography on a ‘thiophilic’ adsorbent. J. Immunol. Meth. 102, 173-182.
- Hutchens, T.W. and Porath, J. (1987). Thiophilic adsorption: A comparison of model protein behavior. Biochem. 26, 7199-7204.
- Hutchens, T.W. and Porath, J. (1987). Protein recognition of immobilized ligands: Promotion of selective adsorption. Clin. Chem. 3319, 1502-1508.
- Oscarsson, S. and Porath, J. (1989). Covalent chromatography and salt-promoted thiophilic adsorption. Anal. Biochem. 176, 330-337.
- Nopper, B., Kohen, F. and Wilchek, M. (1989). A thiophilic adsorbent for the one-step high-performance liquid chromatography purification of monoclonal antibodies. Anal. Biochem. 180, 66-71.
- Lihme, A. and Heegaard, P.M.H. (1990). Thiophilic adsorption chromatography: The separation of serum proteins. Anal. Biochem. 192, 64-69.
- Oscarsson, S., Chaga, G. and Porath, J. (1991). Thiophilic adsorbents for RIA and ELISA procedures. J. Immunol. Meth. 143, 143-149.
Related Resources:
Review of antibody purification methods
Related Products:
Melon Gel Monoclonal IgG Purification Kit (for culture supernatant and ascites)
Melon Gel IgG Purification Kit (for serum samples)
Protein A, Protein G and other IgG purification resins
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