Product Information

Contents: TrueBlot^TM anti-Rabbit Ig IP Beads
Catalog Number: 00-8800
Sizes: 2.5 ml
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
DO NOT FREEZE.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
00-8800	TrueBlot™ anti-Rabbit Ig IP Beads (Binds 2.5mg Ig/ml beads)	N/A	N/A	IP

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

TrueBlot^TM anti-Rabbit Ig IP 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.

Applications Reported

For research use only, not for diagnostic or therapeutic use. TrueBlot^TM anti-Rabbit Ig IP Beads (binds 2.5 mg Ig/ml beads) have been reported for use in immunoprecipitation.

Applications Tested

TrueBlot™ anti-Rabbit Ig IP Beads have been tested by immunoprecipitation and immunoblotting detection using Rabbit TrueBlot™ (cat# 18-8816).

Procedure: Preparation of Immunoprecipitated Sample for SDS-PAGE

1. Preclear cell lysate: Add 50 μl of Anti-Rabbit 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-Rabbit 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 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 100 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 10000xg 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 10000xg 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.

Special Notes

Upon initial use of this product, we recommend that the vial be inverted several times to get the beads into suspension. We recommend to use 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.

Related Products

Cat. 18-8816 Rabbit TrueBlot™: Horseradish Peroxidase (HRP) anti-rabbit IgG