mouse
Articlecode: 00-8811-25
Contents: TrueBlotTM anti-Mouse Ig IP Beads
Catalog Number: 00-8811
Sizes: 2.5 ml
Formulation: 0.5 M NaCl, 0.01% thimerisal
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
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Available Formats of This Product
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Reported Applications
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00-8811
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N/A
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Description
TrueBlot™ anti-Mouse Ig IP Beads are a suspension of activated agarose beads which have been coupled to 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.
Usage
For research use only, not for diagnostic or therapeutic use. TrueBlotTM anti-Mouse Ig IP Beads (binds 0.4 mg Ig/ml beads) have been reported for use in immunoprecipitation.
Applications Tested
TrueBlotTM anti-Mouse Ig IP Beads have been tested by immunoprecipitation and immunoblotting detection using Mouse TrueBlot™ (cat# 18-8877).
Procedure: Preparation of Immunoprecipitated Sample for SDS-PAGE
1. Preclear cell lysate: Add 50 ìl of Anti-Mouse IgG Beads and 500 ìl 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 ìg of primary antibody to the microcentrifuge tube containing the precleared lysate. Incubate on ice for 1 hour. Add 50 ìl of Anti-Mouse IgG Beads. Incubate for 1 hour on a rocking platform. Spin the microcentrifuge tube at 10000xg for 1 minute Remove supernatant completely and wash the (pelleted) beads 3 times with 500 ìl 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 100 ìl Laemmli Buffer (with 50 mM DTT or 2% â-mercaptoethanol, final) to bead pellet. Vortex and heat to 90-100°C for 10 minutes. Spin at 10000xg 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 10000xg 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.
Related Products
Cat. 18-8817 Mouse TrueBlot™ ULTRA: Horseradish Peroxidase (HRP) anti-mouse IgG
Cat. 18-8877 Mouse TrueBlot™: Horseradish Peroxidase (HRP) anti-mouse IgG
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