Magnosphere MS300/Carboxyl particles are magnetic microparticles specially designed for bioseparation such as enzyme immunoassay, IP-western blot or nucleic acid hybridization. The surface of Magnosphere MS300/Carboxyl particles are covered with hydrophilic polymer and carboxyl (-COOH) group, which enable both low non-specific binding and high binding affinity of the probe molecules immobilized on the surface. Probe molecules such as antibodies, proteins and nucleic acids can be immobilized on the surface via amino coupling method shown mannual (See Recommended protocols).
Magnosphere MS300/Low Carboxyl particles are magnetic microparticles specially designed for ultra pure bioseparation prior to high sensitive measurements such as LC-MS.
Nucleic acid hybridization
Example of use 1
Immunoprecipitation of 20S proteasome complex from Jurkat cell lysate
Anti 20S proteasome a6 monoclonal antibody (Biomol International,L.P, Clone MCP20) was conjugated onto the Magnosphere MS300/ Carboxyl through a protocol I.
By using the particle, subunits(a1-a7,b1-b7) of 20S proteasome complex were extracted in a high purity from a Jurkat cell lysate (30mg protein). While, from the parallel experiment using competitor’s magnetic particles, many unexpected noise bands were obtained, but it was difficult to identify bands from prey protein (i.e. proteasome).
|Lane 1 Jurkat cell lysate 4mg protein/lane
Lane 2 IP product using Magnosphere MS300/ Low Carboxyl
Lane 3 IP product using Magnosphere MS300/ Carboxyl
Lane 4 IP product using competitor’s magnetic particles (2.8mm)
Lane 5 IP product using competitor’s magnetic particles (1mm)
Lane 6 IP product using competitor’s magnetic particles (2.8mm)
Lane 7 Molecular weight marker
Antibody conjugated Magnosphere MS300/Low Carboxyl : 0.5mg
Sample : Jurkat cell lysate 30L (100g protein)
IP reaction time : 60 minute (at 4°C)
Washing : Wash 3 times with 0.5mL of 20mM HEPES(PH7.9) + 10v/v%
glycerol + 0.5M KCl + 0.1% NP-40 + 0.1mM EDTA and additional
one time with 0.5mL of TBS-T
Elution : Gentle shaking for 10 minutes at room temperature with 20L
of 0.5% SDS.
Detection : SDS-PAGE, Silver stain
Example of use 2
Sandwich ELISA for Thyroid stimulating hormone (TSH)
As a capture antibody, anti-TSH monoclonal antibody (HyTest, Ltd, Clone 10C7) was conjugated onto the Magnosphere MS300/ Carboxyl through a protocol II. A 50mg of antibody conjugated particle was reacted with TSH spiked human serum for 30 minutes at 37C. The particle was then reacted with secondary antibody labeled with ALP (HyTest, Ltd., Clone:5E8) and chemiluminescence intensity was measured using AMPPD as substrate after washing.
Noise level, signal intensity at 0mIU/mL, from Magnosphere MS300/Carboxyl was about half against the particle using competitor’s and signal intensity at 200mIU/mL was 30% higher than that of competitor’s.
As a result, Magnosphere MS300/Carboxyl showed about 2.5 times higher signal to noise ratio (S/N) than competitor’s particles.
Magnosphere MS300/Low Carboxyl
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.