Language
TrueBlot® Anti-Rabbit Ig IP Agarose Bead
Goat Polyclonal TrueBlot® Anti-Rabbit Ig IP Agarose Bead Properties Description TrueBlot® Anti-Rabbit Ig IP Agarose Beads - 00-8800-25 Host Goat Target Species Rabbit Known Cross Reactivity Rabbit Clonality Polyclonal Application Western Blot : Use with Rabbit TrueBlot® (cat # 18-8816-33) ImmunoPrecipitation: TrueBlot anti-Rabbit Ig IP Beads (binds 2.5 mg Ig/ml beads) have been reported for use in IP Other Dilution: This product is sold individually or in kit p/n 88-1688-31. Species of Origin Goat Product Type Component Kit Type Immunoprecipitation Kit Physical State Suspension of agarose beads Shipping Condition Wet Ice Format IgG Label Agarose beads for TrueBlot® IP/WB Concentration antibody per cc agarose Buffer 0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2 Preservative 0.09% (w/v) Sodium Azide Stabilizer None Relevant Links TrueBlot Protocol | Download Datasheet Download CofA: Lot# |
| |
TrueBlot® Anti-Rabbit Ig IP Agarose Bead Description
Background TrueBlot® Anti-Rabbit Ig IP Agarose Beads are a suspension of activated agarose beads coupled with goat Anti-rabbit IgG. It is suitable for precipitation of rabbit 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. Synonyms Anti-Rabbit Immunoglobulin Gamma, Agarose-conjugated IgG, Gt-a-Rb IgG, Goat-anti-Rabbit IgG, TrueBlot for immunoprecipitation, IP beads for TrueBlot, Anti-Rabbit IgG IP agarose Storage Condition Store vial at 4 °C prior to opening. DO NOT FREEZE. 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-Rabbit IgG Beads and 500μl 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-Rabbit 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-Rabbit 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-Rabbit Ig Beads and immediately transfer an aliquot of the supernatant to gel wells. Disclaimer Note-General 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. 
