ProteinChip H50 Array (Reverse Phase) ® Instruction Manual Catalog #C57-30065 For technical support, call your local Bio-Rad office, or in the US, call 1-800-4BIORAD (1-800-424-6723).
Uses n Protein profiling and biomarker discovery n Rapid protein analysis to determine purity, mass confirmation, or both How It Works The ProteinChip H50 array surface binds proteins through reversephase or hydrophobic interaction chromatography and has binding characteristics similar to that of a C6 to C12 alkyl chromatographic resin. In reverse-phase interactions, proteins within the sample are partitioned between the lipophilic phase of the array surface and the sample buffer.
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. Fig. 2. Removal of ProteinChip arrays from cassette using array forceps.
Recommended Binding and Washing Solutions n ProteinChip H50 buffer (catalog #K20-00001) (10% acetonitrile, 0.1% trifluoroacetic acid (TFA)) n 0–50% methanol or acetonitrile ± 0.1–1% TFA n 0.1% TFA may be added to the binding solution to increase binding Fractionation for Serum Profiling Fractionation of serum prior to profiling is recommended. The fractionation procedure produces 6 fractions containing proteins separated on the basis of their isoelectric point.
Protocol 1: Serum Profiling Using the Bioprocessor Note: These protocols are intended as a guideline; you may need to optimize the method for your particular sample type and experimental design. Note: This protocol is for the 8-spot array in the ProteinChip cassette-compatible bioprocessor. For processing a single array, use the ProteinChip 8-well bioprocessor (catalog #C50-30008). 1.
Protocol 2: Serum Profiling On-Spot 1. Optional: Bulk-wash the ProteinChip array with 50% methanol or acetonitrile for five minutes. Repeat once. Dry the array for an hour after bulk wash to minimize any spot-to-spot cross-contamination. 2. Prewet the spots with 5 µl of binding buffer for 2 minutes. Repeat once. 3. Remove the prewetting solution and replace with 5 µl of sample. Do not allow the spot to air-dry during sample application. 4.
Ordering Information Catalog # Description C57-30065 C50-30011 ProteinChip H50 Arrays, A–H format, 12 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 Prote
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. Bio-Rad Laboratories, Inc. Web site www.bio-rad.
