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Automation Protocol for FLAG Immuno-Precipitation using Magnetic Agarose Beads

 

Product No. M8823

Transfer beads to Binding Plate - NUNC 2000 µl
Step 1: FLAG IP
FLAG magnetic beads (50 µl of a 50% slurry)
  • Plate: SamplePlate, NUNC 2 ml deepwell plate.
  • Incubation time: 4 hrs
  • Agitation: Yes

 

Note: To prevent the accumulation of beads at the bottom of wells, can be separated in 24 x 10 minutes steps with bead collection between each step.

 


Transfer beads to Wash Plate 1 - NUNC 2000 µl
Step 2: Wash 1
  • Volume: 500 µl
  • Incubation time: 5 min
  • Agitation: Yes
 

 


Transfer beads to Wash Plate 2 - NUNC 2000 µl
Step 3: Wash 2
  • Volume: 500 µl
  • Incubation time: 5 min
  • Agitation: Yes
 

 


Transfer beads to Wash Plate 3 - NUNC 2000 µl
Step 4: Wash 3
  • Volume: 500 µl
  • Incubation time: 5 min
  • Agitation: Yes
 

 


Transfer beads to Elution Plate 2 - NUNC 300 µl
Step 5: Elution 1
  • Volume: 50 µl
  • Incubation time: 30 min
  • Agitation: Yes
 

 


Transfer beads to Elution Plate 2 - NUNC 300 µl
Step 6: Elution 1
  • Volume: 50 µl
  • Incubation time: 30 min
  • Agitation: Yes
 

 


IP and Wash Buffer composition:
  • 150 mM NaCl
  • 50 mM Tris pH 7.5
  • 1 mM EDTA
  • 0.5% NP40
  • 10% Glycerol
 

 


Elution Buffer composition: (as above with lower NP40 concentration)
  • 150 mM NaCl
  • 50 mM Tris pH 7.5
  • 1 mM EDTA
  • 0.05% NP40
  • 10% Glycerol
  • FLAG peptide : 500 µg /ml

 



Materials

     
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