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Cat. No.: PAGFI-10 (for 10ml)

Cat. No.: PAGFI-50 (for 50ml)

Description

Protein A+G Agarose (Fast Flow, for IP) is primarily used for immunoprecipitation (IP) or co-immunoprecipitation (Co-IP), and it can also be used for antibody purification.

Protein A is a cell wall surface protein found in Staphylococcus aureus with a molecular weight of 42 kDa. Protein G is an immunoglobulin-binding protein expressed by C-type or G-type Streptococcal bacteria. Protein A and Protein G have similar functions and can specifically bind to mammalian immunoglobulins (Ig). The binding site is usually the Fc region of the immunoglobulin, but there is evidence that Protein A can also bind to the Fab region of the human VH3 family, and Protein G sometimes has a certain affinity for the Fab region as well. Moreover, both proteins have different binding capacities for various subclasses of immunoglobulins. Recombinant Protein A and Protein G, appropriately modified, are combined with agarose gel in a specific manner, making them suitable for immunoprecipitation or antibody purification.

Protein A+G Agarose is suitable for immunoprecipitating all antibodies that can be individually immunoprecipitated by Protein A Agarose and Protein G Agarose, including human IgG1, IgG2, IgG3, IgG4; mouse IgG1, IgG2a, IgG2b, IgG3; rat IgG1, IgG2a, IgG2b, IgG2c; rabbit IgG; and rabbit and goat polyclonal antibodies.

Recombinant Protein A and Protein G in this product can specifically bind to the Fc region of most mammalian IgG antibodies. The molecular weights of Protein A and Protein G are 14 kDa and 22 kDa, respectively. Through modification, these recombinant proteins retain only the amino acid sequences relevant to binding to the Fc region of IgG and remove sequences that may cause non-specific binding outside the binding sites, thereby effectively reducing non-specific binding. Each Protein A molecule and Protein G molecule can bind to 2 and 3 IgG molecules, respectively.

The recombinant Protein A and Protein G in this product are covalently coupled to a 4% cross-linked, high-flow agarose gel (4% Agarose, Fast Flow) and mixed in a 1:1 ratio. Each milliliter of Protein A+G agarose beads (sediment) is coupled with approximately 10 mg of recombinant Protein A and 1 mg of Protein G. Each milliliter of Protein A+G agarose beads (sediment) can bind to over 25 mg of human IgG. The average diameter of the agarose beads in this product is 90 μm. The recommended linear flow rate for antibody purification is 50-300 cm/h, and the maximum pressure tolerance is 0.3 MPa.

This product is formulated in an appropriate solution that can be directly used for immunoprecipitation and antibody purification. Each milliliter contains 0.25 ml of agarose beads (sediment).

If used for routine immunoprecipitation, each milliliter of this Protein A+G Agarose (Fast Flow, for IP) product (a mixture of sediment and solution in a 1:3 ratio) can be used for immunoprecipitation 50 times.

Precautions

• Do not freeze this product for storage.
• Before using Protein A+G Agarose, it must be thoroughly resuspended by gently inverting several times to ensure a uniform mixture.
• This product contains trace amounts of preservatives that do not interfere with routine antibody purification and immunoprecipitation. However, if subsequent enzymatic activity assays are involved, it is recommended to wash the Protein A+G Agarose beads three times with an appropriate solution such as TBS before use to eliminate any potential interference from the preservatives.
• Starting from the collection of protein samples, all steps involving protein samples must be performed at 4°C or on ice.
• This product is intended for scientific research use by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in ordinary households.
• For your safety and health, please wear lab attire and disposable gloves when handling this product.

Storage

Store at 4°C. Valid for one year. Do not freeze.

Only for research and not intended for treatment of humans or animals

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