Protein A/G Magnetic Beads for IP

Protein A/G Magnetic Beads for IP use a biological nanosurface technology (S-TEC). Protein A/G is orientated as a coat on the surface of super paramagnetic microspheres with high coating density up to 9.3 × 10¹³ molecules/cm². Compared to other similar immune magnetic beads, Selleck MagBeads™ Protein A/G display more antibody binding sites, therefore during IP, less magnetic beads are used. Non-specific binding is low, enabling Selleck MagBeads Protein A/G to be used in IP conveniently and efficiently. With a large, specific surface area, these beads can greatly shorten the equilibrium antibody and antigen adsorption time, enabling complete antibody antigen adsorption process within 10 minutes, and complete total purification and precipitation in just 30 minutes. This product can be used on a wide variety of samples, such as in cell lysates, supernatants collected from cell secretion, serum, ascites, and other immune antigens.

Specificity

Protein A/G magnetic beads are affinity purification magnetic beads prepared by separately coupling recombinant Protein A and recombinant Protein G to the surface of magnetic beads. Protein A contains four IgG Fc binding regions, while Protein G possesses two IgG Fc binding regions. Protein A/G magnetic beads combine the advantages of both Protein A and Protein G, enabling efficient capture and purification of a wide range of IgG antibodies from various species and subclasses. Compared to using Protein A or Protein G individually, Protein A/G magnetic beads exhibit stronger IgG binding capacity and are less affected by pH fluctuations during the binding process.

Properties

Concentration	10 mg/mL
Particle size	100 nm
Binding capacity of human IgG	0.4-0.5 mg/mL
Ligand	Recombinant protein A/G
Application	Protein Purification, Immunoprecipitation
pH stability	6-8 (Long term)

This procedure (Figure 1) offers a general guideline for immunoprecipitation (IP). Optimization may be required for each antibody and target antigen. Protein A/G Magnetic Beads for IP are ideally suited for IP reactions.

Figure 1 General Protocol for Immunoprecipitation

Recommended buffer examples

Table 2. Recommended buffer examples

Buffer	Contents (Prepared by customers)
Binding buffer	50 mM Tris, 150 mM NaCl, 0.1%-0.5% detergent (TritonX-100, Tween 20 or NP40), pH 7.5
Wash buffer	50 mM Tris, 150 mM NaCl, 0.1%-0.5% detergent, pH 7.5
Elution buffer	0.1 M -0.2 M Glycine, 0.1%-0.5% detergent, pH 2.5-3.1 (or 0.1 M citric acid, 0.1%-0.5% detergent, pH 2.5-3.1)
Neutralize buffer	1M Tris, pH8.0

Important Notes before Beginning

1. Before immunoprecipitation, please be sure to carefully read the operating instructions.

2. This product requires use of a magnetic separator.

3. Protein A/G Magnetic Beads should be suspended uniformly before use.

4. Protein A/G Magnetic Beads should be kept in storage solution and prevent dry.

5. Do not freeze or centrifuge MagBeads protein A/G.

6. In order to ensure the best results, please select an antibody with strong specificity.

7. For the IP experiments, different antibodies and antigens will display different binding affinities. Antibody and antigen binding may be altered based on use of binding buffer and washing buffer. Some operator optimization may be necessary.

8. This product is only intended to be used as directed. All other uses are prohibited.

Table 1. Relative Binding Strengths of Antibodies to Protein A and Protein G

Species	Antibody class	sProtein A/G	sProtein A
Human	Total IgG	＋＋＋＋＋	＋＋＋＋＋
	IgG1, IgG2	＋＋＋＋＋	＋＋＋＋＋
	IgG3	＋＋＋＋＋	＋
	IgG4	＋＋＋＋＋	＋＋＋＋＋
	IgM	＋	＋
	IgD	－	－
	IgA	＋	＋
	IgA1, IgA2	＋	＋
	IgE	＋＋＋	＋＋＋
	Fab	＋	＋
	ScFv	＋	＋
Mouse	Total IgG	＋＋＋＋＋	＋＋＋＋＋
	IgM	－	－
	IgG1	＋＋＋	＋
	IgG2a	＋＋＋	＋＋＋
	IgG2b	＋＋＋	＋＋＋
	IgG3	＋＋＋	＋＋＋＋＋
Rat	Total IgG	＋＋＋	＋
	IgG 1	＋＋＋	＋＋＋
	IgG2a	＋＋＋＋＋	＋＋＋
	IgG2b	＋	＋＋＋
	IgG2c	＋＋＋＋＋	＋＋＋
Cow	Total IgG	＋＋＋＋＋	＋
	IgG1	＋＋＋＋＋	＋
	IgG2	＋＋＋＋＋	＋＋＋＋＋
Goat	Total IgG	＋＋＋＋＋	＋
	IgG1	＋＋＋＋＋	＋
	IgG2	＋＋＋＋＋	＋＋＋＋＋
Sheep	Total IgG	＋＋＋＋＋	＋
	IgG1	＋＋＋＋＋	＋
	IgG2	＋＋＋＋＋	＋＋＋＋＋
Horse	Total IgG	＋＋＋＋＋	＋
	IgG(ab), IgG(c)	＋	－
	IgG(T)	＋＋＋＋＋	＋
Rabbit	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Guinea pig	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Hamster	Total IgG	＋＋＋	＋＋＋
Pig	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Donkey	Total IgG	＋＋＋＋＋	＋＋＋
Cat	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Dog	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Monkey	Total IgG	＋＋＋＋＋	＋＋＋＋＋
Chicken	Total IgG	－	－

Notes: "+"= weak binding, "+++"=medium binding, "+++++"=strong binding, "-"=no binding

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