NHS Activated MagBeads


MagBeads
PureCube NHS Activated MagBeads carry a N-hydroxy-succinimide (NHS) functionality. Proteins can bind covalently via amine groups, forming a peptide bond. Our careful in-house production provides low lot-to-lot variation, making activated MagBeads also suitable for commercial production. PureCube NHS Activated MagBeads are delivered as a 25% suspension.

PureCube NHS Activated MagBeads provide:


Features

Usage Covalent coupling of biomolecules via amine groups
Specifity Affinity to free amine groups of e.g. proteins
Binding capacity 20 mg / mL
Carboxylic group densits higher than 15 µmol/ml
Filling quantity Delivered as a 25 % suspension
Bead size 25 μm
Bead Ligand Activated NHS
Required equipment
 

Applications

All protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.NHS coupling protocol
 
 
  1. Transfer 1 mL PureCube NHS-Activated MagBeads 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 beads once with 1 ml PBS. Allow the beads to separate and remove the supernatant. Important: Once PBS is added, work quickly to avoid hydrolysis of the NHS groups.
  4. Prepare a solution of 625 µL PBS containg the protein to be coupled to the MagBeads. The exact protein amount needs to be optimized, and 1 to 3 mg protein is a good starting point. 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 MagBeads and mix by vortexing.
  6. Depending on the temperature stability of the protein, incubate at room temperature or 4°C for 2 h on an end-overend shaker or thermoshaker.
  7. Place the tube on a magnetic stand and allow the beads to separate. 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. Add 1.5 mL PBS buffer to the MagBeads, mix by vortexing, and separate on a magnetic stand. Remove the supernatant.
  9. Repeat step 7.
  10. Wash four times with 1.5 mL double distilled water each.
  11. Add 1.2 mL Quenching Buffer and incubate again for 1 h at room temperature or for 4 hours at 4°C.
  12. Wash four times with 1.5 mL PBS each, and twice with 1.5 mL double distilled water each.
  13. Resuspend the coupled MagBeads in 1 mL MagBead Storage buffer, yielding a 25% suspension. Store at 4°C.
MagBeads
PureCube NHS Activated MagBeads carry a N-hydroxy-succinimide (NHS) functionality. Proteins can bind covalently via amine groups, forming a peptide bond. Our careful in-house production provides low lot-to-lot variation, making activated MagBeads also suitable for commercial production. PureCube NHS Activated MagBeads are delivered as a 25% suspension.

PureCube NHS Activated MagBeads provide:


Features

Usage Covalent coupling of biomolecules via amine groups
Specifity Affinity to free amine groups of e.g. proteins
Binding capacity 20 mg / mL
Carboxylic group densits higher than 15 µmol/ml
Filling quantity Delivered as a 25 % suspension
Bead size 25 μm
Bead Ligand Activated NHS
Required equipment
 

Applications

All protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.NHS coupling protocol
 
 
  1. Transfer 1 mL PureCube NHS-Activated MagBeads 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 beads once with 1 ml PBS. Allow the beads to separate and remove the supernatant. Important: Once PBS is added, work quickly to avoid hydrolysis of the NHS groups.
  4. Prepare a solution of 625 µL PBS containg the protein to be coupled to the MagBeads. The exact protein amount needs to be optimized, and 1 to 3 mg protein is a good starting point. 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 MagBeads and mix by vortexing.
  6. Depending on the temperature stability of the protein, incubate at room temperature or 4°C for 2 h on an end-overend shaker or thermoshaker.
  7. Place the tube on a magnetic stand and allow the beads to separate. 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. Add 1.5 mL PBS buffer to the MagBeads, mix by vortexing, and separate on a magnetic stand. Remove the supernatant.
  9. Repeat step 7.
  10. Wash four times with 1.5 mL double distilled water each.
  11. Add 1.2 mL Quenching Buffer and incubate again for 1 h at room temperature or for 4 hours at 4°C.
  12. Wash four times with 1.5 mL PBS each, and twice with 1.5 mL double distilled water each.
  13. Resuspend the coupled MagBeads in 1 mL MagBead Storage buffer, yielding a 25% suspension. Store at 4°C.
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NHS Activated MagBeads
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PureCube NHS Activated MagBeads PureCube NHS Activated MagBeads
Protein purification ► NHS-Activated Magnetic Beads, for direct coupling of biomolecules via amine groups. ★25% conc.★
Article number: 50401
Sales price: From €107.00 *
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