ProteinChip®NP20 Array
(Normal Phase)
Instruction Manual
Catalog #C57-30043
For technical support,
call your local Bio-Rad office, or
in the US, call 1-800-4BIORAD
(1-800-424-6723).
Uses
n
Binding of highly water-soluble proteins
n
On-chip sample cleanup
n
Calibration of the ProteinChip SELDI system instrument using
peptides or proteins of known molecular weight
How It Works
Silicon oxide groups on the active spots of the ProteinChip NP20
array allow proteins to bind through hydrophilic and charged
residues, including serine, threonine, and lysine. This surface is
commonly used for microscale sample cleanup. Washing with
water removes buffer components such as salts and detergents.
Packaging and Storage
Store the arrays at room temperature.
ProteinChip arrays are packaged in a 12-array cassette.
A bioprocessor reservoir is included in the package (see Figure 1).
The spare ProteinChip cassette included to separate the reservoir
from the arrays should be removed before use in the ProteinChip
cassette-compatible bioprocessor (catalog #C50-30011). It is not
necessary to remove the arrays when using the cassette-compatible
bioprocessor; however, individual arrays can be removed if needed.
To do this, remove the bioprocessor reservoir before taking any
arrays out of the cassette. Be careful not to touch the spots on the
array. A pair of ProteinChip array forceps (catalog #C20-10002)
helps effectively remove the arrays from the cassette (see Figure 2).
Fig. 1. ProteinChip cassette and
reservoir.
© 2006 Bio-Rad Laboratories, Inc.
Fig. 2. Removal of ProteinChip arrays
from cassette using array forceps.
Technical Considerations
n
Prerinsing the ProteinChip NP20 array with HPLC or better
grade water may improve results
Protocol: On-Spot
Note: This protocol is intended as a guideline; you may need to optimize the method
for your particular sample type and experimental design.
1. Dilute or dissolve the sample in binding buffer. Most buffers are
tolerated by this surface.
2. Load 5 µl of sample per spot (ideally 200–2,000 nM, 10-fold less
for peptides).
3. Allow sample to air-dry onto the surface for 5–10 minutes.
4. Wash each spot with 5 µl of binding buffer with shaking
(on MicroMix shaker, setting 20/4) and remove buffer.
Repeat two more times.
5. Air-dry the array for 15–20 minutes.
6. Apply 0.5 µl of saturated ProteinChip energy absorbing
molecules (EAM) in solution to each spot. Allow to air-dry.
Optional: Add another 0.5 µl of saturated EAM in solution to
improve signal intensity.
7. Analyze the arrays using the ProteinChip SELDI system.
© 2006 Bio-Rad Laboratories, Inc.
Life Science
Group
06-0669 1006 Sig 1205
10008233 US/EG Rev H
Bio-Rad
Laboratories, Inc.
Web site www.bio-rad.com USA 800 4BIORAD
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Ordering Information
atalog # Description
C
57-30043
C
C50-30011
C50-30008
C50-30012
C20-10002
C30-00001
C30-00002
C30-00003
MicroMix is a trademark of Diagnostic Products Corporation.
The SELDI process is covered by US patents 5,719,060, 6,225,047, 6,579,719,
6,818,411, and other issued patents and pending applications in the US and other
jurisdictions.
roteinChip NP20 Arrays,A–H format, 12
P
ProteinChip Cassette-Compatible Bioprocessor, includes
ProteinChip array forceps, cassette hold-down frame, 12 blank
ProteinChip arrays
ProteinChip 8-Well Bioprocessor, A–H format
ProteinChip Cassette-Compatible Bioprocessor Reservoirs, 5
ProteinChip Array Forceps, 1 pair
ProteinChip CHCA Energy Absorbing Molecules (EAM),
5 mg/vial, 20
ProteinChip SPA Energy Absorbing Molecules (EAM),
5 mg/vial, 20
ProteinChip EAM-1 Energy Absorbing Molecules (EAM),
5 mg/vial, 20
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