Contents: TrueBlot™ anti-Goat Ig IP Beads Catalog Number: 00-8844 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
00-8844	TrueBlot™ anti-Goat Ig IP Beads (Binds 1.0mg Ig/ml beads)	N/A	N/A	IP

TrueBlot™ anti-Goat Ig IP Beads have been tested by immunoprecipitation and immunoblotting detection using Goat TrueBlot™ (cat# 18-8814).

Procedure: Preparation of Immunoprecipitated Sample for SDS-PAGE

1. Preclear cell lysate: Add 50 ìl of anti-goat IgG beads and 500 ìl of cell lysate sample to an eppendorf tube and incubate on ice for 30 minutes. Spin at 10,000xg for 3 minutes and transfer the supernatant to a new eppendorf tube.
2. Immunoprecipitation: Add 5 ìg of primary antibody to the eppendorf tube containing the precleared lysate. Incubate on ice for 1 hour. Add 50 ìl of Anti-Goat IgG Beads. Incubate for 1 hour on a rocking platform. Spin the eppendorf tube at 10000xg for 1 minute. Remove supernatant completely and wash the (pelleted) beads 3 times with 500 ìl of Lysis Buffer.
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) or NuPAGE LDS Sample Buffer (Invitrogen Cat#NP0007) containing NuPAGE Sample Reducing Agent (dithiothreitol; Invitrogen Cat#NP0004) 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-goat Ig beads.
Note: The supernatant can be stored at -20°C for future use. After thawing, add fresh NuPAGE Sample Reducing Agent (dithiothreitol) and heat as above. Centrifuge the sample at 10000xg for 1 minute in a microcentrifuge to pellet any anti-goat Ig beads and immediately transfer an aliquot of the supernatant to gel wells.