FLAG Immuno-Precipitation Protocol using Magnetic Agarose Beads

Product No. M8823

Transfer beads to Binding Plate - NUNC 2000 µl

Step 1: FLAG IP

FLAG magnetic beads (50 ml 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

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