SweAgarose Protein A/G Antibody Purified Agarose

Protein A is a cell wall surface protein found in Staphylococcus aureus with a molecular weight of 42kDa. Protein G is an immunoglobulin-binding protein expressed by Streptococcal bacteria (C or G).Protein A and Protein G are functionally similar and can bind specifically to mammalian immunoglobulin (Ig). Recombinant Protein A and G with appropriate modifications bound to agarose can be used for immunoprecipitation or antibody purification. Protein A agarose are suitable for the immunoprecipitation of human IgG1, IgG2, IgG4, mouse IgG2a, IgG2b, while Protein G agarose are suitable for the immunoprecipitation of human IgG1, IgG2, IgG3, IgG4, mouse IgG1, IgG2a, IgG2b, IgG3, rat IgG1, IgG2a, IgG2b, IgG2c polyclonal antibodies. (See Table I for specific information)

This product adopts the self-developed and produced Protein A/G labelled agarose, compared with similar products in the market, this product binds antibodies more efficiently and has a lower non-specific binding rate, together with the optimized buffer, it can be convenient and efficient for immunoprecipitation experiments; it can be widely used in the isolation and purification of the target proteins in samples such as cell lysates, cellular secretion supernatants, serum, ascites, etc.

Ship with wet ice; Store at 2-8℃, valid for 12 months.

Antibody purification related reagent formulations refer to the following, the user can be adjusted according to specific experimental conditions.

Manual procedure (purification of mouse ascites IgG as an example)

1. Sample processing: take 500 μL of ascites sample, add the binding wash buffer to make up 500 μL, if there are more protein precipitates in the sample, centrifuge the supernatant for experiments, which can improve the purity of the antibody.

2. Agarose pretreatment: Vortex the antibody purified agarose for 30 s to resuspend sufficiently, take 100 μL of 50% (v/v) SweAgarose Protein A/G Antibody Purified agarose in another new 1.5 mL EP tube, centrifuge at 6,000 x g for 30s at 4℃ and discard the supernatant, wash it with 1 mL of binding washing buffer for twice, and then take the supernatant after magnetically aspirating.

Note: The amount of agarose can be adjusted according to the amount of antibody in the sample.

3. Antibody adsorption: add the sample processed in step 1 to the agarose in step 2, vortex and mix well, place the EP tube in a rotary mixer or manually turn the tube gently at room temperature (about 25°C) to make the agarose full contact with the sample, turn it over for 15 min, centrifuge at 6,000 x g for 30s at 4℃, and then discard the supernatant.

4. Agarose washing: add 1 mL of binding wash buffer to the EP tube, resuspend with shaking and then centrifuge at 6,000 x g for 30s at 4℃. Discard the supernatant and repeat the operation 3 times.

5. Antibody elution: Add 0.5~1.0 mL of eluent to the EP tubes with the agarose washed as described above, and resuspend the tubes rapidly by pipetting or vortexing, and then gently turn the tubes over in a turnover mixer or by hand at room temperature (about 25℃), centrifuge at 6,000 x g for 30s at 4℃ after turning over for 10 min, and then collect the supernatant into new EP tubes.

6. Antibody neutralisation: add a certain amount of neutralisation solution to the antibody eluate in step 5, generally 1/10 of the antibody elution volume (e.g., if the antibody eluate is 500 μL, the amount of neutralisation solution added is 50 μL), so that the pH value of the eluted antibody is maintained in a neutral environment, which is conducive to the maintenance of the biological activity of the antibody and the avoid antibody inactivation.

7. Post-treatment of Agarose: Wash the agarose twice with elution solution after use, centrifuge at 6,000 x g for 30s at 4℃, and discard the supernatant; then wash three times with binding washing solution, centrifuge at 6,000 x g for 30s at 4℃, and discard the supernatant; resuspend the agarose with 200 µL of preservation solution, and then store at 2~8℃.

1. Please read these instructions carefully before proceeding with antibody purification.

2. Do not freeze or centrifuge the agarose as this may cause irreversible aggregation of the agarose.

Table 1

For research use only. Not for use in diagnostic or therapeutic procedures!