Instruction Manual

ANNEX A: CELLSEARCH SAMPLE
PREPARATION FOR DEPArray
Prepare the CELLSEARCH sample for DEPArray with VR NxT
Materials required
Name Volume Provider Code
Corning Thick Gel Loading Pipet
Tip
1-200 μl Round 0.5 mm
Corning 4853
Protein LoBind tube
1.5 mL
Eppendorf
l EU:0030108.116
l US: 022431081
Research plus pipette 20-200 μl Eppendorf P200 pipette: 3120000054
100-1000 μl P1000 pipette: 3120000062
epT.I.P.S LoRetention ep Dualfilter 20-300 μl Eppendorf l EU: 0030077.636
l US: 022493004
50-1000 μl l EU: 0030077.652
l US: 022493008
Swing bucket upstream centrifuge for 1.5 mL tube Any supplier -
Bovine Serum Albumin, lyophilized
powder (BSA)
- Any supplier -
1X PBS - Any supplier -
DEPArray Buffer for fixed cells - Menarini Silicon
Biosystems
KI0066
Getting started
1. Thaw a bottle of DABUF and keep it at room temperature.
NOTICE: Discard any unused DABUF once thawed. Do not refreeze it.
2. Prepare a BSA 2% solution in 1X PBS (for example, 1 g of BSA in 50 mL of 1X PBS), considering that the minimal
volume required per sample is 1.5 mL.
3. Let the solution sit and then gently agitate and invert it to mix.
NOTICE: Do not vortex.
4. Prepare the required set of 1.5 mL tubes per sample.
Required set of 1.5 mL tubes per sample
Tube quantity Content of the tubes Name to write on the tube
2 325 μl of DABUF each DABUF
1 1.5 mL of 2% BSA in 1X PBS BSA
1 empty, to collect the sample later Sample
1 empty, to collect Supernatant 1 later S1
1 empty, to collect Supernatant 2 later S2
1 empty, to keep the gel loading tip after BSA coating Tip
Coat the pipette tip with the BSA
1. Set the P200 pipette to 200 μl.
2. Attach a clean gel loading tip to the P200 pipette.
3. Immerse the gel loading tip in the BSA tube since the internal and external surfaces of the tip are coated.
4. Slowly pipette up and down the BSA solution five times.
5. Discard any solution and bubbles present in the tip.
VR NxT| Instruction Manual 1.0 02-2019|VRNXTinstr1EN|© 2019 Menarini Silicon Biosystems SpA 45