Affinity Ligand Coated Microspheres

Affinity binding ligands can aid in the rapid development or streamlined production of microsphere reagents. They also provide the opportunity for optimal orientation of the ligand, as with end-point immobilization of biotinylated oligonucleotides to streptavidin-coated microspheres, or the Fc-specific immobilization of antibodies via Protein A or G.

Affinity ligands are also used extensively in isolation and purifications. Streptavidin-coated microspheres are often used to capture biotinylated PCR amplicons. Protein A and Protein G microspheres are utilized for immunoprecipitation and the purification of antibodies and Fc-tagged fusion proteins. Lectin-coated spheres (e.g. WGA, ConA) are used to bind glycoproteins, glycolipids and other glycan-presenting targets.

Our streptavidin and SuperAvidin™ coated microspheres offer simple and extremely stable binding of biotinylated molecules, such as antibodies, peptides, and oligonucleotides for use in diagnostics, screening, and affinity purifications. See TechNote 101, Affinity Ligand coated microspheres. Streptavidin coated microspheres are available as ~1% solids (w/v) aqueous suspensions in these standard volumes: 1mL, 2mL, 5mL, or 10mL. See also our BioMag® magnetic particle offerings.

Catalog Number Nominal Diameter Specification Range
CP01000 0.10µm 0.090 – 0.110µm
CP01001 0.20µm 0.190 – 0.210µm
CP01003 0.50µm 0.470 – 0.530µm
CP01004 1.00µm 0.95 – 1.05µm
CP01005 3.0µm 2.80 – 3.20µm
CP01006 5.00µm 4.80 – 5.20µm
CP01007 10.00µm 9.50 – 10.50µm
CP01008 15.0µm 14.50 – 15.50µm

REFERENCES

  • Nandin IS, Fong C, Deantonio C, Torreno-Pina J, Pecetta S, Maldonado P, … Batista, F.(2017) Novel in vitro booster vaccination to rapidly generate antigen-specific human monoclonal antibodies. J Exp Med.20170633. (0.11µm Streptavidin-coated microspheres)
  • Wu SJ,Chan A, Kado CI. (2004) Detection of PCR amplicons from bacterial pathogens using microsphere agglutination J Microbiol Methods Vol 56, Issue 3, 395-400 (5.7µm Streptavidin-coated microspheres)
  • Walter S, HerrgenL, Schoor O, Jung G, Wernet D, Bühring HJ, … Stevanović S.(2003) Cutting Edge: Predetermined Avidity of Human CD8 T Cells Expanded on Calibrated MHC/Anti-CD28-Coated Microspheres. J Immunol. 171(10)4974-4978 (5.6µm Streptavidin as artificial-antigen presenting cells [aAPC])
  • Jeffers, Faye, et al. “Mucin–lectin interactions assessed by flow cytometry.” Carbohydrate research 345.10 (2010): 1486-1491.(3.09µm Streptavidin PS & 2.01µm Silica)

The surfaces of Encapsulated magnetic microspheres are expected to be free of iron oxide, while the Classical version will have some exposed iron oxide. COMPEL microspheres (UMC catalog codes) are highly uniform and spherical. Fluorescent magnetic microspheres are available as ~1% solids (w/v) aqueous suspensions in these standard volumes: 1mL, 5mL, 10mL, and 100mL. See spectra for available fluorescent dyes. See also our BioMag® magnetic particle offerings.

*Please note: If you are NOT ordering via website, please specify a lot number if multiple lots are available.
Catalog Number Description Nominal Diameter Specification Range
CME0101 SA / Encapsulated Magnetic Polymer 0.86µm 0.7 – 1.2µm
UMC0100 SA / COMPEL 3µm 2.50 – 3.50µm
UMC0101 SA / COMPEL 6µm 5.50 – 6.50µm
UMC0102 SA / COMPEL 8µm 7.50 – 8.50µm

REFERENCES

  • Pitsillides, CM, Joe, EK, Wei, X, Anderson, RR, & Lin, CP. (2003). Selective cell targeting with light-absorbing microparticles and nanoparticles. Biophysical journal, 84(6), 4023-4032.(0.83µm Streptavidin magnetic)
Catalog Number Description
MFYS1N Magnefy™ • SA 1µm
PMS1N ProMag™ 1 Series • Streptavidin
PMS3HP ProMag™ HP 3 Series • Streptavidin
PMS3N ProMag™ 3 Series • Streptavidin
21950 Streptavidin Magnetic Sample Pack MFYS1N, Magnefy 1µm SA – 1mL (1% solids) PMS1N, ProMag 1 Series SA – 1mL (1% solids) PMS3HP, ProMag HP 3μm SA – 1mL (1% solids) BP628, BioMagPlus SA – 2mL (5mg / mL)

References

  • Linz TH, Henley WH, & Ramsey JM. (2017) Photobleaching kinetics-based bead encoding for multiplexed bioassays. Lab on a Chip, 17(6), 1076-1082. (3µm ProMag Streptavidin with immobilized b-PE and b-Qdot labels)
Catalog Number Description Concentration (mg/mL) Volume (mL)
BM551 BioMag® Streptavidin 5 5, 50
BP628 BioMag®Plus Streptavidin 5 10
BM568 BioMag® Streptavidin Nuclease-free 1 10, 25
BP621 BioMag®Plus Streptavidin/Biotin Binding Kit 5 5

REFERENCES

  • Chang D, Tram K, Li B, Feng Q, Shen Z, Lee CH, … & Li, Y. (2017). Detection of DNA Amplicons of Polymerase Chain Reaction Using Litmus Test. Scientific reports, 7(1), 3110.(Cat. BM551BioMag Streptavidin used in colorimetric detection of C. difficile)
Catalog Number Fluorescence Color Nominal Diameter Specification Range
CFDG001 SA / Dragon Green 0.20µm 0.190 – 0.210µm
CFDG002 SA / Dragon Green 0.40µm 0.370 – 0.430µm
CFFR002 SA / Flash Red 0.40µm 0.370 – 0.430µm
CFDG003 SA / Dragon Green 0.50µm 0.470 – 0.530µm
CFDG004 SA / Dragon Green 1.00µm 0.95 – 1.05µm
CFFR004 SA / Flash Red 1.00µm 0.95 – 1.05µm

REFERENCES

  • Bhattacharyya, K., Goldschmidt, B. S., & Viator, J. A. (2016). Detection and capture of breast cancer cells with photoacoustic flow cytometry. Journal of biomedical optics, 21(8), 087007. (320nm streptavidin-coated Flash Red fluorescent nanoparticles + biotin-EpCAM)

Streptavidin-coated dyed microspheres supplied as ~1% solids (w/v) aqueous suspensions.

Catalog Number Coating / Dye Nominal Diameter Specification Range
CDCR001 SA / Crimson Red 0.20µm 0.190 – 0.210µm
CDCB001 SA / Cabo Blue 0.20µm 0.190 – 0.210µm

References

  • Wang Y, Yan W, Fu S, Hu S, Wang Y, Xu J, & Ye C. (2018). Multiple cross displacement amplification coupled with nanoparticles-based lateral flow biosensor for detection of Staphylococcus aureus and identification of methicillin-resistant S. aureus. Frontiers in Microbiology, 9.(0.129µm Streptavidin-coated Crimson Red nanoparticles)

SiO2 with a streptavidin-coated surface for the attachment of biomolecules (See also PDS 721.pdf). Silica streptavidin microspheres are sold in these standard amounts: 1mL, 2mL, 5mL, and 10mL. Available at 1% solids (w/v), in three nominal mean diameters: ~0.5µm, ~1.0µm, and ~5.0µm.

Catalog Number Nominal Diameter Specification Range
CS01000 0.5µm 0.460 – 0.540µm
CS01001 1.0µm 0.95 – 1.05µm
CS01002 5.00µm 4.80 – 5.20µm

REFERENCES

  • Blumberg, S, Gajraj, A, Pennington, M, Meiners, J (2005) Three-Dimensional Characterization of Tethered Microspheres by Total Internal Reflection Fluorescence Microscopy. Biophysical Journal V89(2)p1272–1281
  • Wang J, Xu D, Kawde A, Polsky R (2001) Metal Nanoparticle-Based Electrochemical Stripping Potentiometric Detection of DNA Hybridization Anal Chem. 73(22), 5576-5581

Our biotin-coated microspheres have been fully characterized in terms of their ability to bind free avidin, and therefore will require minimal optimization when determining the correct concentration of ligand to be bound. Biotin coated beads are supplied as ~1% solids (w/v) aqueous suspensions. See TechNote 101, Affinity Ligand coated microspheres, for sample applications and procedures. See also our BioMag® offerings.

Catalog Number Description Nominal Diameter Specification Range
CP10000 Polymeric / Biotin 10µm 9.50 – 10.50µm

Protein A and Protein G are proteins that readily bind the Fc regions of many types of antibodies. (See TSD 0025 ) For this reason, they are used extensively for affinity purification of antibodies from serum or cell culture supernatant, and for capture of immunoprecipitated protein complexes. When Protein A and Protein G microspheres are used as reagents in tests and assays, the immobilized antibodies may be crosslinked to the Protein A- or Protein G-coated support for long-term stability. See TechNote 101, Affinity Ligand coated microspheres. Protein A and Protein G coated microspheres are available as ~1% solids (w/v) aqueous suspensions in these standard volumes: 1mL, 2mL, 5mL, or 10mL. See also our BioMag® offerings.

Catalog Number Description Nominal Diameter Specification Range
CP02000 Polymer / Protein A 1.00µm 0.95-1.05µm
CP02001 Polymer / Protein A 5.00µm 4.80-5.20µm
CP02002 Polymer / Protein A 10.0µm 9.50-10.50µm
PMG3N ProMag™ (Magnetic) / Protein G 3µm 2.70 – 3.30µm

REFERENCES

  • Reynolds NM, Mohammadalipour A, Hall CR, Adib AA, Farnoud AM, Burdick MM. (2018) Galectin-1 Influences Breast Cancer Cell Adhesion to E-selectin Via Ligand Intermediaries. Cell Mol Bioeng; 11(1):37-52. Note:  (Protein A / E-selectin [hFc] beads used to investigate E-selectin / Gal-1 mediated adhesion in breast cancer monolayers)

BioMag® secondary antibody particles are used extensively in the capture of immune complexes for immunoprecipitation and antibody-labeled cells for indirect cell separation. Secondary antibody coatings also afford a convenient method for binding specific primary antibodies to BioMag® particles.

Catalog Number Description Concentration (mg / mL) Volume (mL)
BM563 BioMag®  Goat anti-Human IgG 1 50
BM562 BioMag® Goat anti-Human IgG (Fc Specific) 5 50
BM561 BioMag® Goat anti-Human IgM 1 50
BM549 BioMag® Goat anti-Mouse IgG 1 50 & 500
BM550 BioMag® Goat anti-Mouse IgG (Fc Specific) 1 50 & 500
BP619 BioMag®Plus Goat anti-Mouse IgG 1 50
BP612 BioMag®Plus Goat anti-Mouse IgG Antibody Coupling Kit 1 25
BM558 BioMag® Goat anti-Mouse IgM 1 50 & 500
BM559 BioMag® Goat anti-Rabbit IgG 1 50 & 500
BM560 BioMag® Goat anti-Rat IgG 1 50 & 500
BM548 BioMag® Goat anti-Rat IgG (Fc specific) 5 50 & 500
BM557 BioMag® Goat anti-Rat IgM 5 50 & 500

Our ConA- and WGA coated BioMag®Plus microparticles provide a convenient means for isolating glycoproteins from serum or cell lysate, and for investigating other lectin/glycan-mediated processes. The BioMag®Plus magnetic particle format provides high surface area, and permits easy and efficient separations.

Catalog Number Description Concentration (mg/mL) Volume (mL)
BP530 BioMag®Plus Wheat Germ Agglutinin 5 3, 10
BP531 BioMag® Plus Concanavalin A 5 3, 10

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