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Cat. No.: PAFI-10 (for 10ml)

Cat. No.: PAFI-50 (for 50ml)

Description

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

Protein A is a cell wall surface protein found in Staphylococcus aureus bacteria 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 certain binding to the Fab region as well. Additionally, both Protein A and Protein G have different binding capacities for different subclasses of immunoglobulins. Recombinant modified Protein A and Protein G are coupled to agarose beads in a specific manner, making them suitable for immunoprecipitation or antibody purification.

Protein A Agarose is suitable for immunoprecipitating human IgG1, IgG2, IgG4; mouse IgG2a, IgG2b; and rabbit IgG, among others.

Recombinant Protein A in this product specifically binds to the Fc region of most mammalian IgG molecules, with a molecular weight of 14 kDa. This recombinant Protein A has been genetically engineered to become alkali-resistant by introducing specific modifications, while retaining only the amino acid sequence relevant for binding to the IgG Fc region. This modification eliminates non-specific binding sequences outside the binding site, effectively reducing non-specific binding. Each Protein A molecule can bind to 2 IgG molecules.

The recombinant Protein A in this product is covalently coupled to 4% cross-linked agarose beads with high flow rate (4% Agarose, Fast Flow). Each milliliter of Protein A agarose beads (sediment) is coupled with approximately 20 mg of recombinant Protein A. Each milliliter of Protein A agarose beads (sediment) can bind to over 30 mg of human IgG. The leaching of Protein A is extremely low, with less than 15 ng of leached Protein A per milligram of purified IgG. The average diameter of the agarose beads in this product is 90 μm, and the recommended linear flow rate for antibody purification is 50-300 cm/h, with a maximum pressure tolerance of 0.3 MPa. It operates in a pH range of 3-12 and can withstand 0.1-0.5 M NaOH.

This product is prepared in a suitable solution that can be directly used for immunoprecipitation and antibody purification, with a total volume of 0.25 ml of agarose beads per milliliter.

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

Precautions

• Do not freeze this product for storage.
• Before using Protein A 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 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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