Maleimide Activated MagBeads


MagBeads
PureCube Maleimide Activated MagBeads available as off-the-shelf items. Our careful in-house production provides low lot-to-lot variation, making activated MagBeads also suitable for commercial production. Maleimide activated MagBeads are provided as a 25% suspension. PureCube Maleimide Activated MagBeads carry a maleimide functionality. Proteins can bind covalently and irreversibly to free thiol groups.

PureCube Maleimide Activated MagBeads provide:

  • Covalent, irreversible binding via free thiol groups
  • Pre-activated matrix for coupling to proteins, peptides, or antibodies
  • Easy to follow protocols and expert technical support
  • Also available as Maleimide Activated Agarose

Maleimide activated MagBeads from Cube Biotech were successfully used in the following publications and patents:

Purified SubstanceYearAuthor
DNA 2019 Toldrà A., Alcaraz C., Diogène J., O'Sullivan C.K., Campàs M.1
P2-GM1os 2019 Mahon C.S., Wildsmith G. C., Haksar D., de Poel E., Beekman J.M., Pieters R.J., Webb M.E., Turnbill W.B.,2
Affimers, A11, B7, and G10 2018 Klont F., Hadderingh M., Horvatovich P., ten hacken N.H.T., Bischoff R.,3
Tetrodotoxins 2018 Leonardo S., Kiparissis S., Rambla-Alegre M., Almarza S., Roque A., Andree K.B., Christidis A., Flores C., Caixach J., Campbell K., Elliot C.T., Aligizaki K., Diogène J., Campàs M.4
Cystein-SpyCatcher-Protein 2018 Pröschel M.,5
DNA 2018 Lamble H.J., Egan C., Lloyd D., Dunski E.,6

Features

Usage Purification Or Immobilization of proteins or DNA
Coupling mechanism Coupling to Maleimide via thiother bonds
Filling quantity Delivered as a 25 % suspension
Bead Ligand Maleimide
Required equipment
 
  • Sodium dihydrogen phosphate
  • Sodium chloride
  • Sodium acetate trihydrate
  • Sodium hydroxide (NaOH)
  • Acetic acid
  • Ethanol
  • Optional: Sodium hydrogen carbonate
  • Optional: Optional: Sodium azide
  • Optional: Tris(2-chlorethyl)phosphate (TCEP)
  • Magnetic holder for microcentrifuge tubes (for separation of magnetic beads)
  •  Microcentrifuge tubes (2 mL)
  •  pH meter
  • Vortex
  • End-over-end shaker
  • SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment

Applications

   
A.Maleimide coupling protocol:
 
 
  1. Transfer 1 mL PureCube Maleimide-Activated MagBeads (25%) into a 2mL microcentrifuge tube. Tip: The coupling reaction can be linearly scaled up and down, by increasing or decreasing the amounts of buffers and solutions described in this protocol.
  2. Place the tube on a magnetic stand and allow the beads to separate. Remove the supernatant.
  3. Wash the Activated MagBeads three times with 1 ml coupling buffer I or II. Tip: Choose coupling buffer I or II depending on the pH stability of your protein. Use the same coupling buffer throughout the procedure.
  4. Resuspend 0.5-1.5 mg protein in 250 µl coupling buffer I or II. Tip: When coupling a particular protein for the first time, try 3-5 different protein concentrations to make sure you are offering enough protein in the reaction but not wasting any protein.
  5. Add the protein solution to the magnetic beads and mix by vortexing.
  6. Depending on the temperature stability of the protein, incubate at 20°C for 2 hours or at 4° C overnight on an endover-end shaker or thermoshaker.
  7. Remove the supernatant and analyze the supernatant in a spectrophotometer. Record absorption at 280 nm to monitor coupling efficiency. Tip: Monitoring absorbance at 280 nm tells you about the coupling efficiency of the protein (compare A280 of the original protein solution to the supernatant in step 6 to determine % coupling). It also helps you identify the optimal amount of protein required for efficient coupling.
  8. Wash the magbeads three times with 1 ml coupling buffer and once with 1 ml double distilled water.
  9. Resuspend the coupled MagBeads in 1 mL Storage buffer I or II, yielding a 25% suspension. Store at 4°C.

References

1. Toldrà, A. et al. (2019). Detection of Ostreopsis cf. ovata in environmental samples using an electrochemical DNA-based biosensor. Science of The Total Environment. doi:10.1016/j.scitotenv.2019.06.448
2. Mahon, C. et al. (2019). A “catch-and-release” receptor for the cholera toxin. Faraday Discussions. doi:10.1039/c9fd00017h
3. Klont, F. 1t al.(2018). Affimers as an Alternative to Antibodies in an Affinity LC–MS Assay for Quantification of the Soluble Receptor of Advanced Glycation End-Products (sRAGE) in Human Serum. Journal of Proteome Research, 17(8), 2892–2899. doi:10.1021/acs.jproteome.8b00414
4. Leonardo, S. et al. (2019). Detection of tetrodotoxins in juvenile pufferfish Lagocephalus sceleratus (Gmelin, 1789) from the North Aegean Sea (Greece) by an electrochemical magnetic bead-based immunosensing tool. Food Chemistry, 290, 255–262. doi:10.1016/j.foodchem.2019.03.148
5. Pröschel M. Identifizierung, Charakterisierung und Optimierung von neuen kovalenten Split-Isopeptid Systemen. Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU). https://opus4.kobv.de/opus4-fau/frontdoor/index/index/docId/9621
6. Lamble H.J., Egan C., Lloyd D., Dunski E., inventor; Sense Biodetection Limited, assignee. Nucleid Acid Detection Method. United States patent US2018/0216157. 2018 August 2.
MagBeads
PureCube Maleimide Activated MagBeads available as off-the-shelf items. Our careful in-house production provides low lot-to-lot variation, making activated MagBeads also suitable for commercial production. Maleimide activated MagBeads are provided as a 25% suspension. PureCube Maleimide Activated MagBeads carry a maleimide functionality. Proteins can bind covalently and irreversibly to free thiol groups.

PureCube Maleimide Activated MagBeads provide:

  • Covalent, irreversible binding via free thiol groups
  • Pre-activated matrix for coupling to proteins, peptides, or antibodies
  • Easy to follow protocols and expert technical support
  • Also available as Maleimide Activated Agarose

Maleimide activated MagBeads from Cube Biotech were successfully used in the following publications and patents:

Purified SubstanceYearAuthor
DNA 2019 Toldrà A., Alcaraz C., Diogène J., O'Sullivan C.K., Campàs M.1
P2-GM1os 2019 Mahon C.S., Wildsmith G. C., Haksar D., de Poel E., Beekman J.M., Pieters R.J., Webb M.E., Turnbill W.B.,2
Affimers, A11, B7, and G10 2018 Klont F., Hadderingh M., Horvatovich P., ten hacken N.H.T., Bischoff R.,3
Tetrodotoxins 2018 Leonardo S., Kiparissis S., Rambla-Alegre M., Almarza S., Roque A., Andree K.B., Christidis A., Flores C., Caixach J., Campbell K., Elliot C.T., Aligizaki K., Diogène J., Campàs M.4
Cystein-SpyCatcher-Protein 2018 Pröschel M.,5
DNA 2018 Lamble H.J., Egan C., Lloyd D., Dunski E.,6

Features

Usage Purification Or Immobilization of proteins or DNA
Coupling mechanism Coupling to Maleimide via thiother bonds
Filling quantity Delivered as a 25 % suspension
Bead Ligand Maleimide
Required equipment
 
  • Sodium dihydrogen phosphate
  • Sodium chloride
  • Sodium acetate trihydrate
  • Sodium hydroxide (NaOH)
  • Acetic acid
  • Ethanol
  • Optional: Sodium hydrogen carbonate
  • Optional: Optional: Sodium azide
  • Optional: Tris(2-chlorethyl)phosphate (TCEP)
  • Magnetic holder for microcentrifuge tubes (for separation of magnetic beads)
  •  Microcentrifuge tubes (2 mL)
  •  pH meter
  • Vortex
  • End-over-end shaker
  • SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment

Applications

   
A.Maleimide coupling protocol:
 
 
  1. Transfer 1 mL PureCube Maleimide-Activated MagBeads (25%) into a 2mL microcentrifuge tube. Tip: The coupling reaction can be linearly scaled up and down, by increasing or decreasing the amounts of buffers and solutions described in this protocol.
  2. Place the tube on a magnetic stand and allow the beads to separate. Remove the supernatant.
  3. Wash the Activated MagBeads three times with 1 ml coupling buffer I or II. Tip: Choose coupling buffer I or II depending on the pH stability of your protein. Use the same coupling buffer throughout the procedure.
  4. Resuspend 0.5-1.5 mg protein in 250 µl coupling buffer I or II. Tip: When coupling a particular protein for the first time, try 3-5 different protein concentrations to make sure you are offering enough protein in the reaction but not wasting any protein.
  5. Add the protein solution to the magnetic beads and mix by vortexing.
  6. Depending on the temperature stability of the protein, incubate at 20°C for 2 hours or at 4° C overnight on an endover-end shaker or thermoshaker.
  7. Remove the supernatant and analyze the supernatant in a spectrophotometer. Record absorption at 280 nm to monitor coupling efficiency. Tip: Monitoring absorbance at 280 nm tells you about the coupling efficiency of the protein (compare A280 of the original protein solution to the supernatant in step 6 to determine % coupling). It also helps you identify the optimal amount of protein required for efficient coupling.
  8. Wash the magbeads three times with 1 ml coupling buffer and once with 1 ml double distilled water.
  9. Resuspend the coupled MagBeads in 1 mL Storage buffer I or II, yielding a 25% suspension. Store at 4°C.

References

1. Toldrà, A. et al. (2019). Detection of Ostreopsis cf. ovata in environmental samples using an electrochemical DNA-based biosensor. Science of The Total Environment. doi:10.1016/j.scitotenv.2019.06.448
2. Mahon, C. et al. (2019). A “catch-and-release” receptor for the cholera toxin. Faraday Discussions. doi:10.1039/c9fd00017h
3. Klont, F. 1t al.(2018). Affimers as an Alternative to Antibodies in an Affinity LC–MS Assay for Quantification of the Soluble Receptor of Advanced Glycation End-Products (sRAGE) in Human Serum. Journal of Proteome Research, 17(8), 2892–2899. doi:10.1021/acs.jproteome.8b00414
4. Leonardo, S. et al. (2019). Detection of tetrodotoxins in juvenile pufferfish Lagocephalus sceleratus (Gmelin, 1789) from the North Aegean Sea (Greece) by an electrochemical magnetic bead-based immunosensing tool. Food Chemistry, 290, 255–262. doi:10.1016/j.foodchem.2019.03.148
5. Pröschel M. Identifizierung, Charakterisierung und Optimierung von neuen kovalenten Split-Isopeptid Systemen. Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU). https://opus4.kobv.de/opus4-fau/frontdoor/index/index/docId/9621
6. Lamble H.J., Egan C., Lloyd D., Dunski E., inventor; Sense Biodetection Limited, assignee. Nucleid Acid Detection Method. United States patent US2018/0216157. 2018 August 2.
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Maleimide Activated MagBeads
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PureCube Maleimide Activated MagBeads PureCube Maleimide Activated MagBeads
1 ml 25% Maleimide-Activated MagBeads, for direct, irreversible coupling of biomolecules via thiol groups  
Article number: 51201
Sales price: From €135.00 *
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