It is not difficult to immobilize a protein on a solid support using any classic activation method such as the cyanogen bromide or N-hydroxysuccinimidylcarboxylate... However, if the protein of interest is a specific IgG, its biological activity will be affected if not entirely lost after the covalent linkage between the solid support with the terminal H₂N of the IgG.

AFFILAND has developed the Activated Protein A-Sepharose fast flow useful for the immobilization of IgG on the F_c portion, leaving free the Fab: the biological properties of the specific IgG will be conserved.

CHARACTERISTICS OF THE GEL

Matrix: Sepharose^TM 4 Fast Flow
Ligand: Activated Protein A which has a specific binding for the F_c of IgG. It does not recognize serum albumin or serum proteins other than immunoglobulins.
Rapid formation of a stable complex between the ligand and the F_c portion of immunoglobulin from IgG from mouse, rabbit, human..., in the binding & coupling buffer, followed by an immediate covalent linkage between the Activated matrix and the F_c portion, leaving free the F_ab of IgG.
Easily realized in a column.
Sample: can be either the antisera, ascites fluid, extract containing antibodies or purified IgG.
Stability of the matrix: pH 6-9.
Storage: 4°C in PBS 50mM pH 7.4; NaN₃ 0.02% (w/v).

Results

Easy recovery of the antibody immobilized on the gel in a record time of 30 minutes maximum. The capacity of the obtained affinity matrix may be 5-6 mg of mouse monoclonal IgG or 10-12 mg of polyclonal IgG of most other animal species, per ml of matrix.

APPLICATIONS

• Purification of antigens by immunoaffinity chromatography.
• Easy preparation of serum free of antigen.
• Radio or enzyme immunoassay.

ORDERING INFORMATION

Product code		Product name	Prepackaged column	*Price in EUR
APA-M2 KIT	APA-M2	Activated Protein A - Sepharose fast flow column (for APA-M2 KIT)	3x1 ml gel column	230.00
	BPA-M2	Binding & Coupling buffer (for APA-M2 KIT)	500 ml
	SRP-M2	Saturation reagent (for APA-M2 KIT)	50 ml

* All prices are FOB Liège.
Bulk quantities are available.

INSTRUCTIONS FOR USE WILL BE PROVIDED WITH THE KIT.