Anti-GFP Magnetic Beads

Anti-GFP Magnetic Beads
Product Description 
Anti- GFP magnetic beads are based on carboxyl magnetic beads, with 200 nm particle size, covalently coupling with high quality mouse IgG monoclonal antibody that recognizes the GFP tag (Green Fluorenscent Protein). 
Product Features 
Composition mouse IgG monoclonal Ab Magnetization Superparamagnetic Particle size 200 nm Concentration 10 mg/mL Binding Capacity ≥ 0.6 mg GFP-tagged fusion protei n/mL of bead Application IP，CoIP Storage Condition Store at 4°C for 2 years. 
Protocol 
1. Cell lysis Cells may be lysed using any standard cell lysis protocol compatible with your starting material. We recommend the use of Cell Extraction Buffer or NP-40 Cell Lysis Buffer. 2. Preparation of Magnetic Beads 2.1   Resuspend the Magnetic Beads in the vial (tilt and rotate for 2 minutes or gently pipette for 10 times). 2.2   Transfer 10-20 μL of Anti- GFP Magnetic Beads into a 1.5 mL tube (Transfer amount may be adjusted as required). 2.3   Add 500 μL of binding/wash buffer to the beads and gently pipette to mix. Place the tube into a magnetic stand to collect the beads against the side of the tube (Hereinafter referred to as magnetic separation). Remove and discard the supernatant. Repeat this step for 2 times. 3. Immunoprecipitation  3.1 Remove the tubes from the magnetic separator and add your sample containing GFP-tagged protein to the pre-washed magnetic beads and incubate at room temperature for 2h or overnight at 4°C with mixing. 3.2 Collect the beads with a magnetic stand, remove the unbound sample and save for analysis. 
 
 
 
3.3 Add 500μL of TBST to the tube and gently mix. Collect the beads and discard the supernatant. Repeat this wash twice. 3.4 Add 500μL of ultrapure water to the tube and gently mix. Collect the beads on a magnetic stand and discard the supernatant. 4. Elution Chemical Elution Protocols • Basic Elution 1. Add 100μL of 1x SDS-PAGE loading buffer to the tube. 2. Boil for 5 minutes on a dry bath. 3. Magnetically separate the beads and save the supernatant containing the target antigen. • Acidic Elution 1. Add 100μL of 0.1M glycine, pH 3.0. 2. Gently vortex to mix and incubate the sample at room temperature on a rotator for 5-10 minutes. 3. Magnetically separate the beads and save the supernatant containing the target antigen. 4. To neutralize the low pH, add 15μL of Neutralization Buffer (1 M Tris pH 8.0) for each 100μL of eluate. The final solution can be used as samples for denaturing SDS-PAGE. Or the elution can be adjusted to neutral pH with neutralization buffer immediately and used for further analysis.