ExalphaÕs IP-Readyª agarose suspension products contain 0.5 ml of agarose conjugate and are ready to use for immunoprecipitation procedures. They are provided preblocked with fatty acid free bovine serum albumin and have been extensively washed. Formulations are ready to use, 10 µl of agarose per 1 µg of primary antibody used in standard immunoprecipitation procedures.
Supplied 0.25 ml of preblocked Protein A agarose and 0.25 ml of preblocked Protein G agarose in 1 ml of phosphate buffered saline containing 0.08% sodium azide.
Antigen-antibody binding interaction
Protein A/G Agarose IP-Ready antibody Antibody Purification Reagent
Antibody's reacts with
Antibody's suited for
Basic procedure for immunoprecipitation assay using IP-Readyª products. These are basic guidelines and may need to be modified for your particular application:_x000B__x000B_Prep of PBSTDS: Mix 100 ml of 10x PBS with 10 ml of 100% Triton X-100; 5 g of sodium deoxycholate and 1.0 g sodium dodecyl sulfate. Include a cocktail of protease inhibitors, such as 1 mM EDTA; 1 µg/ml Pepstatin and 0.2 mM PMSF from 100 mM stock in 2-propanol, or a cocktail of you choosing; Dilute this solution to 1 L with distilled water._x000B__x000B_1. Preclear lysate or media by adding ~1 µg of normal IgG from the same species from which the primary antibody is derived to remove nonspecific proteins._x000B_2. Incubate for 1 hour at 4¼C and then centrifuge at 1,500 rpm reserving supernatant for specific IP._x000B_3. Determine total protein of the sample._x000B_4. To a 1.5 ml Eppendorf tube, add the following: Cell lysate (1 mg total protein), 1 µg of purified primary antibody and 10 µl of packed beads._x000B_5. Cap tubes and incubate at 4¼C on rocker platform or rotating device for 2 to 24 hours._x000B_6. Centrifuge at 2,500 rpm for 15 minutes at 4oC and collect immunoprecipitate._x000B_7. Wash pellet 4 times with 1.0 ml of PBSTDS, repeating centrifuge step above (step 6) each time._x000B_8. After final wash, aspirate and discard supernatant and resuspend pellet in 40-50 µl of electrophoresis sample buffer._x000B_9. Analyze samples by either SDS-PAGE or transfer to a membrane and detect by Western blot.
The purest agarose was used in the production of Protein A/G Agarose IP- antibody Antibody Purification Reagent by nordc.If you buy Antibodies supplied by nordc they should be stored frozen at - 24°C for long term storage and for short term at + 5°C.
Antibodies are affinity purified with an antigen coated column or protein A or G agarose or beads. DNA is purified with endotoxin free silica columns or anion exchange resins. nordc supplies purification kits and ultra pure reagents.
Protein A/G Agarose IP- antibody Antibody Purification Reagent
Purification, Antibody, Agarose, IP
Protein A/G molecular sieve IP-Ready (Antibody to) Antibody purification Reagent