Protein A Agarose IP-ReadyªCatalogue number: X1203
|Product Type||Antibody Purification Reagent|
Exalpha's 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.
Synonyms: IP-Ready(tm) Protein A Agarose
Product Form: Agarose
Formulation: Supplied 0.5 ml of preblocked Protein A agarose in 1 ml of phosphate buffered saline containing 0.08% sodium azide.
Basic procedure for immunoprecipitation assay using IP-Ready™ products. These are basic guidelines and may need to be modified for your particular application: 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. 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. 2. Incubate for 1 hour at 4ºC and then centrifuge at 1,500 rpm reserving supernatant for specific IP. 3. Determine total protein of the sample. 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. 5. Cap tubes and incubate at 4ºC on rocker platform or rotating device for 2 to 24 hours. 6. Centrifuge at 2,500 rpm for 15 minutes at 4oC and collect immunoprecipitate. 7. Wash pellet 4 times with 1.0 ml of PBSTDS, repeating centrifuge step above (step 6) each time. 8. After final wash, aspirate and discard supernatant and resuspend pellet in 40-50 µl of electrophoresis sample buffer. 9. Analyze samples by either SDS-PAGE or transfer to a membrane and detect by Western blot.
Product should be stored at 4-8ºC. DO NOT FREEZE
Product Stability: Products are stable for one year from purchase when stored properly
Ship Conditions: Ship at ambient temperature, do not freeze, refrigerate upon arrival
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals.
Safety Datasheet(s) for this product:
Protein A Agarose IP-Readyª