MS Series

MS300/Carboxyl

Magnosphere^TM MS300/Carboxyl is a magnetic particle that enables high-purity bioseparation. Our proprietary particle surface design reduces non-specific binding and maintains the affinity of binding probes which allows the recovery of the target substance at a high S/N ratio. This series can be also used in immunoassays. Try this particle if the target antibody cannot be activated when bound to MK Series or MB Series.

Characteristics

Hydrophilic surface
Low non-specific binding
High probe binding affinity

Specification

Particle size	3μm
Surface carboxy density	10 nmol/mg beads approx.
Solid content	1% (approx. 6x10⁸ particles/ml)
Magnetite content	20% approx.

Protocol

Magnosphere MS300/Carboxyl

Application Example

1. Immunoprecipitation of the 20S proteasome complex from Jurkat cell lysate

The 20S proteasome is a protein which is composed of 28 subunits: 14α-subunits and 14β-subunits that form a four stacked ring. The anti-20S proteasome alpha 6 monoclonal antibody (Biomol International,L.P., clone MCP20) was conjugated to Magnosphere^TM MS300/Carboxyl. The Magnosphere^TM particles with the antibody were then used to immunoprecipitate the 20S proteasome complex from a Jurkat cell lysate (30mg protein). High-purity proteasome complexes (α1 ~ α7, β1 ~ β7) were recovered by Magnosphere^TM MS300/Carboxyl particles.

2. Chemiluminescence enzyme immunoassay (CLEIA) for thyroid-stimulating hormone (TSH)

The anti-TSH monoclonal antibody (HyTest Ltd., clone 10C7) was immobilized on Magnosphere^TM MS300/Carboxyl. Each of the human serum specimens (50 μL) into which the TSH antigen was spiked at different concentrations was reacted with 50 μg of particles for 30 min at 37℃, followed by incubation with an alkaline phosphatase labeled secondary antibody (HyTest Ltd., clone 5E8) labeled by alkaline phosphatase (ALP). A substrate solution (AMPPD) was then added to the specimen, and chemiluminescence was measured.
Compared to the competitor's magnetic particle (2.8 μm), the noise level at an antigen concentration of 0 mIU/mL for Magnosphere^TM MS300/Carboxyl was about half times lower. On the other hand, the signal intensity at an antigen concentration of 200 mIU/mL was 1.3 times higher. A high sensitivity with an S/N ratio of about 2.5 times was obtained.

Antigen Concentration (μIU/mL)	Magnosphere	Competitor's Magnetic Particle
0	80	150
5	40060	27431
60	268542	191409
200	412546	312716

MS300/Low Carboxyl

Magnosphere^TM MS300/Low Carboxyl is a magnetic particle that enables high-purity bioseparation. Magnosphere^TM MS300/Low Carboxyl beads have even lower non-specific binding surface properties compared to Magnosphere^TM MS300/Carboxyl beads .

Characteristics

Hydrophilic surface
Very low non-specific binding
High probe binding affinity

Specification

Particle size	3μm
Surface carboxy density	3 nmol/mg beads approx.
Solid content	1% (approx. 6x10⁸ particles/ml)
Magnetite content	20% approx.

Protocol

Magnosphere MS300/Low Carboxyl

Immunoprecipitation (IP) of the 20S proteasome complex from Jurkat cell lysate

The anti-20S proteasome alpha 6 monoclonal antibody (Biomol International,L.P., clone MCP20) was conjugated to Magnosphere^TM MS300/Carboxyl and Magnosphere^TM MS300/ Low Carboxyl. The Magnosphere particles with the antibody were then used to immunoprecipitate the 20S proteasome complex from a Jurkat cell lysate (30mg protein). High-purity proteasome complexes (α1 ～ α7, β1 ～ β7) were recovered from Magnosphere^TM MS300/Carboxyl and Magnosphere^TM MS300/Low Carboxyl. The noise level was lower in Magnosphere^TM MS300/Low Carboxyl than in Magnosphere^TM MS300/Carboxyl. When three types of magnetic particles from a competitor were used, many non-specific proteins were observed, making the detection of the target substance difficult.

<Operating conditions>
Antibody-bound Magnosphere^TM MS300/Low Carboxyl: 1 mg
Sample: Jurkat cell lysate 100mL (30 mg protein)
Immunoprecipitation response time: 60 min (4℃)
Lavage: lavaged three times (5 sec each) with a 0.5 mL solution of lavage fluid 20 mM HEPES (PH7.9) + 10 v/v% glycerol + 0.5 M KCl + 0.1% NP-40 + 0.1 mM EDTA, and then lavaged once with TBS-T.
Elution: shaken for 10 min at room temperature, with 20 mL 0.5% SDS added.
Detection; SDS-PAGE, silver staining

Lane 1	Jurkat cell lysate 4mg protein/lane
Lane 2	IP product obtained by using MagnosphereTM MS300/Low Carboxyl as the carrier
Lane 3	IP product obtained by using MagnosphereTM MS300/Carboxyl as the carrier
Lane 4	IP product obtained by using the competitor’s magnetic particle (2.8 mm) as the carrier
Lane 5	IP product obtained by using the competitor’s magnetic particle (1 mm) as the carrier
Lane 6	IP product obtained by using the competitor’s magnetic particle (2.8 mm) as the carrier
Lane 7	Molecular weight marker

MS300/Streptavidin

Magnosphere^TM MS300/Streptavidin beads are magnetic microparticles coated with streptavidin for bioseparation. The particle surfaces are covered with a JSR proprietary hydrophilic polymer to give the beads their characteristic low non-specific binding without inhibition of enzyme activities. Consequently, Magnosphere^TM MS300/Streptavidin beads can be used for a variety of applications such as PCR or immunoassays for excellent performance.

Characteristics

High affinity to biotinylated proteins and nucleotides
Hydrophilic surface
Low non-specific binding

Specification

Particle size	3μm
Biotin binding capacity	400-500 pmol/mg beads
Solid content	1% (approx. 6x10⁸ particles/ml)
Magnetite content	20% approx.

More info

Magnosphere MS300/Streptavidin

Application Example

1. Effect on PCR

Quantitative PCR was performed with Magnosphere^TM MS300/Streptavidin added to a known concentration of 100b.p.DNA, and the Threshold Cycle (Ct) was plotted. Ct was constant with no PCR inhibition up to 3 mg of Magnosphere^TM MS300/Streptavidin added to the reaction.

2.Chemiluminescence enzyme immunoassay (CLEIA) for a-fetoprotein (AFP)

An EIA test was performed by binding a biotinylated anti-AFP antibody to Magnosphere^TM MS300/Streptavidin and the competitor's avidin particle. The signal intensity was measured at different AFP concentrations. Magnosphere^TM MS300/Streptavidin showed a higher signal level than the competitor's particle.

Literature

The use of Magnosphere^TM MS300/Tosyl paramagnetic beads in HCV and EBV assay formats

Back to Magnosphere