Natural Protein A is a cell wall surface protein found in Staphylococcus aureus, it is similar to protein G isolated from the  genus  G  or  C  Streptococcus, bind  most  mammalian  IgG  by  interacting  primarily  with  the  Fc  region  of immunoglobulin (Ig), but differ in their binding specificity. It can be used for the separation and purification of a variety of antibodies (monoclonal antibody and polyclonal antibody). Protein A and Protein G differ in their binding properties to different immunoglobulin subclasses (see Appendix 1 for details).
This product uses genetically modified protein A, which not only maintains its Ig affinity properties, but also remove the non-major binding domain of the natural protein itself to reduce non-specific binding. SuRi rProtein A Agarose Resin, which is highly cross-linked agarose gel as the matrix and modified alkali-resistant recombinant protein A as a ligand, has high physicochemical stability. High purity antibody can be obtained from ascites, serum and culture medium samples by one-step affinity chromatography with this product, which is convenient to use and widely used.

Components

Specifications

Shipping and Storage

The products are shipped with ice pack.   It can be stably stored at 2~8℃ for 5 years.

Instructions

1.  Buffer preparation

It is recommended that the following buffers be filtered with a 0.22 μm or 0.45 μm filter membrane before use. Balance/bind/wash buffer: 0.15 M NaCl, 20 mM Na2 HPO4, pH 7.0

Elution buffer: 0.1 M glycine, pH 3.0

Neutralizing buffer: 1 MTris-HCl, pH 8.5

2. Sample preparation

Ensure that the sample solution has the appropriate ionic strength and pH before loading the column, either by diluting the serum sample, ascites, or cell culture solution with a bind/wash buffer, or by dialysis the sample with a bind/wash buffer.

3. Column Packing

Load the SuRi rProtein A Agarose Resin into a suitable chromatographic column, taking care to avoid bubbles.

4. Sample purification

1) Balance: The chromatographic column is balanced with a binding Buffer of 5 times the column volume, so that the packing is in the same buffer system as the target protein, and plays the role of protecting the protein.

2) Sample loading: The sample was loaded to the well-balanced rProtein A Agarose Resin to ensure full contact between the target protein and the resin, improve the recovery rate of the target protein, and collect the effluent to be tested.

3) Washing: Wash with 10-15 times the column volume of the washing Buffer, remove the impurity protein unless specifically adsorbed, collect the washing solution.

4) Elution: Elution with 5-10 times column volume of the elution Buffer, collect eluent, that is, the target protein component.

5) CIP cleaning and preservation: Generally use 0.1-0.5 M NaOH greater than 3CV, and the contact time is 15 min; Then rinse with 5CV balancing solution until neutral. Finally, it is balanced with 20% ethanol of 5 times the column volume,  and  then  stored  in  20%  ethanol  of  the  same  volume  at  4°C  to  prevent  the  packing  from  being contaminated by bacteria.

5. SDS-PAGE detection

The samples obtained in the purification process (including original samples, effluent components, washing and elution components, etc.) were detected by SDS-PAGE to determine the purification effect.

Notes

1.  Do not refrigerate the product.
2.  rProteinAAgarose Resin must be fully reversed several times before use, so that the agarose resin mixed evenly.
3.  During all operations, the sample needs to be operated at 4°C or on ice.
4.  Foryour safety and health, please wear lab coats and disposable gloves for operation.
5.  For research use only.

Attached table 1 : Summary of binding ability of proteins A and G to Ig in different species

SuRi rProtein A Agarose Resin