Application Note: Upon initial use of this product, we recommend that the vial be inverted several times to get the beads into suspension. We recommend using a large bore pipet to pipet up the liquid for use. For storage of the opened vial, we recommend that the vial cap be sealed with parafilm to help prevent evaporation of the buffer. Procedure: Preparation of Immunoprecipitated Sample for SDS-PAGE: 1. Preclear cell lysate: Add 50 μL of Anti-Mouse IgG Beads and 500 μL of cell lysate sample to a microcentrifuge tube and incubate on ice for 30 minutes. Spin at 10,000xg for 3 minutes and transfer the supernatant to a new microcentrifuge tube. 2. Immunoprecipitation: Add 5 μg of primary antibody to the microcentrifuge tube containing the precleared lysate. Incubate on ice for 1 hour. Add 50 μL of Anti-Mouse IgG Beads. Incubate for 1 hour on a rocking platform. Spin the microcentrifuge tube at 10,000xg for 1 minute. Remove supernatant completely and wash the (pelleted) beads 3 times with 500 μL of Lysis Buffer (50mM Tris HCl, pH 8.0; 150mM NaCl; 1% NP-40). 3. Prepare sample for SDS-PAGE: After the last wash, aspirate supernatant, and add 50 μL Laemmli Buffer (with 50 mM DTT or 2% β-mercaptoethanol, final) to bead pellet. Vortex and heat to 90-100 °C for 10 minutes. Spin at 10,000xg for 3 minutes, collect supernatant, and load onto the gel. Avoid loading Anti-Mouse Ig Beads. Note: The supernatant can be stored at -20 °C for future use. After thawing, add fresh dithiothreitol and heat as above. Centrifuge the sample at 10,000xg for 1 minute in a microcentrifuge tube to pellet any Anti-Mouse Ig Beads and immediately transfer an aliquot of the supernatant to gel wells.
Concentration Value: 14.4 mg/cc
Immunoprecipitation Dilution: TrueBlot anti-Mouse Ig IP Beads (binds 0.4 mg Ig/ml beads) have been reported for use in IP
Western Blot Dilution: Use with Mouse TrueBlot® (cat # 18-8817-33)
General Disclaimer Note: This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
Physical State: Suspension of agarose beads
Purity and Specificity: TrueBlot® Anti-Mouse Ig IP Agarose Beads has been tested in SDS-Page, immunoprecipitation, and western blot.
Background: TrueBlot® Anti-Mouse Ig IP Agarose Beads are a suspension of activated agarose beads coupled with goat Anti-mouse IgG. It is suitable for precipitation of mouse IgGs used as the primary antibodies in immunoprecipitation assays. The beads are in suspension and will settle upon storage. Prior to use, mix the vial gently (do not vortex) to ensure delivery of proper bead volume.
Low Endotoxin: No
Other: This product can be sold individually or in kit p/n 88-7788-31.
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