Cat. No.: HY-K0209

Anti-His Magnetic Beads

Name: Anti-His Magnetic Beads
Brand: MedChemExpress
SKU: HY-K0209
Price: 274.00 USD
Availability: InStock
Rating: 5 (27 reviews)

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Based on 27 publication(s) in Google Scholar

Storage :
4°C, 2 years. Do not centrifuge, dry or freeze the magnetic beads.

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Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Description & Advantages

MCE Anti-His Magnetic Beads are used for immunoprecipitation (IP) of specific His-tagged proteins expressed in bacterial and mammalian cells and in vitro expression systems. Anti-His 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 His tag. Magnetic beads are removed from the solution manually by using a magnetic stand or automatically by using an instrument. With high loading of His-tagged protein and high specificity, Anti-His Magnetic Beads are also suitable for Co-immunoprecipitation and purification of His-tagged protein.

Characteristics
Composition	Mouse IgG monoclonal antibody covalently coupled to a blocked magnetic bead surface
Bead Diameter	200 nm
Binding Capacity	> 0.5 mg protein/mL of beads
Application	IP, Co-IP, Protein Purification
Recommended Dose	25 μL for per 500 μL cell lysates

Protocols

1. Preparation of Magnetic Beads

1.1 Resuspend the Magnetic Beads in the vial (tilt and rotate for 2 minutes or gently pipette for 10 times, do not vortex). Transfer 25 μL of Anti-c-Myc Magnetic Beads suspension into a new tube.

1.2 Add 500 μL of wash buffer to the beads and gently pipette to mix. Place the tube into a magnetic stand (MCE Cat. No.: HY-K0200) to collect the beads against the side of the tube. Remove and discard the supernatant. Repeat this step for 2 times

2. Protein Binding

2.1 Add 500 μL of cell lysate (the sample containing c-Myc-tagged protein) to the washed beads. For Ag binding, incubate for 2 hours at room temperature or overnight at 4°C while gently rotating the tube.

2.2 Place the tube into a magnetic stand to collect the beads against the side of the tube. Remove and discard the supernatant.

NOTE: Occasional aggregation of magnetic beads during the binding process doesn’t affect experimental results.

3. Washing

Add 500 μL of wash buffer to the Magbeads-Ag complex and mix gently. Place the tube into a magnetic stand to collect the beads against the side of the tube. Remove and discard the supernatant. Repeat this step for 3 times.

4. Elution & Detection

Three elution methods are recommended according to protein characteristics or further usage:

1) Elution with sample buffer for gel electrophoresis and immuoblotting. Add 100 μL of 1× SDS-PAGE loading buffer to each tube and boil for 5 minutes. Cool and place the tube into a magnetic stand to collect the beads and transfer the supernatant to a new tube. Keep the supernatant containing the target antigen for SDS-PAGE analysis

2) Elution with Elution Buffer A under acidic condition. Add 100 μL of Elution Buffer A to each tube. Incubate with gentle shaking or on a rotator for 10 minutes at room temperature. Place the tube into a magnetic stand to collect the beads and transfer the supernatant to a new tube. Adding 10-20 μL of Neutralization Buffer for each 100 μL of eluate to neutralize the low pH, which may help preserve bioactivity of target protein.

3) Elution with Elution Buffer B under native condition. Add 3-5 (v/v) volume of Elution Buffer B to each tube. Incubate with gentle shaking or on a rotator for 1 hour at room temperature or 2 hours at 4°C. Place the tube into a magnetic stand to collect the beads and transfer the supernatant to a new tube. For immediate use, store the eluates at 4°C, or store at -20°C for long term storage.