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Pack Size | Price | Availability | Quantity |
---|---|---|---|
1 mL | $218 | In Stock | |
5 mL | $858 | In Stock |
Reagent | Formulation |
---|---|
Washing Buffer (1×) | TBST: 50 mM Tris-HCl,150 mM NaCl, 0.1%(v/v) Tween-20,pH7.4 |
Acidity Elution Buffer | 0.1 M Glycine,0.1% (v/v) Tween-20,pH2.5 |
Neutralization Buffer | 1 M Tris-HCl, pH 9.0 |
GST Tag Immunomagnetic Beads | Features |
---|---|
Material | Silica-based magnetic beads |
Bead Size | 200 nm |
Concentration | 10 mg/mL |
Binding Capacity | ≥ 0.6 mg GST tag protein/mL beads |
Ligand | Anti-GST monoclonal antibody produced in mouse |
Recommended Applications | IP, Co-IP |
1. Preparation of cell lysates
Select an appropriate lysis buffer to lyse cell samples and obtain cell lysates. Place on ice or store at -20℃ for long-term use.
2. Pretreatment of Magnetic Beads
(1) Vortex for 1 min to resuspend the immunomagnetic beads. Take 25-50 µL of suspension and place it in a 1.5 mL EP tube.
(2) Add 500 μL of Washing Buffer to the EP tube and gently invert several times to resuspend the beads. Keep the EP tube in a magnetic separator and stand for 1 min for magnetic separation. Finally, remove the supernatant and then take off the EP tube. Repeat the washing steps twice.
3. Immunoprecipitation
(1) Add 500 μL of prepared cell lysates to the EP tube. Place it on a rotating mixer and rotate at 37℃ for 30 min. For weak binding, incubate at room temperature for 1 hour or overnight at 4℃.
(2) After incubation, perform magnetic separation, then remove or save the supernatant for further analysis.
(3) Add 500 μL of Washing Buffer to the EP tube. Perform magnetic separation. Finally, remove the supernatant and then take off the EP tube. Repeat the washing steps 3 times.
4. Elution of Target Proteins
(1) Denaturing Elution: Suitable for SDS-PAGE detection. Add 100 μL of SDS-PAGE Loading Buffer to the EP tube. Mix well and heat at 95℃ for 5 min. Perform magnetic separation or centrifugation (room temperature, 13000 g, 10 min) to collect the supernatant.
(2) Acidity Elution: Add 100 μL of Acidity Elution Buffer to the EP tube. Incubate on a rotating mixer at 37℃ for 5-10 min. Perform magnetic separation or centrifugation to collect the supernatant. To adjust the pH of acidic elution buffer to neutral, add 50 μL of Neutralization Buffer to 100 μL elution.
Store at 4℃ for 2 years.
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