ProteinChip Q10 Array (Strong Anion Exchanger) ® Instruction Manual Catalog #C57-30080 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 operating mechanism of ionic-exchange ProteinChip arrays is the reversible binding of charged molecules to the surface, and the property of a peptide or protein that governs its binding is its net surface charge. Since surface charge is the result of weak acidic and basic amino acids within the protein, binding of the protein to the array is highly pH dependent.
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 cassettecompatible bioprocessor; however, individual arrays can be removed if needed.
Technical Considerations n Increasing the ionic concentration of buffer and/or lowering the pH of the binding and washing buffers will increase the selectivity of the surface n If salts are present in the sample (>50 mM), remove with spin column n Choose a binding buffer that can buffer at least one pH unit above the isoelectric point (pI) of the target protein(s) n When using a bioprocessor, make sure there are no air bubbles in the wells.
2. Remove the buffer from the wells. Immediately add 50–150 µl sample to each well. Recommended concentration is 50–2,000 µg/ml total protein, diluted in binding buffer. Incubate with vigorous shaking for 30 minutes. 3. Remove the samples from the wells and wash each well with 150–250 µl binding buffer for 5 minutes, with agitation. Repeat two more times. 4. Remove the binding buffer from the wells and add 150–250 µl deionized (DI) water to each well, remove immediately. Repeat once. 5.
Protocol 2: Serum Profiling On-Spot 1. Prewet the spots with 5 µl of binding buffer for 5 minutes. Repeat once. 2. Remove the prewetting solution and replace with 5 µl of sample. Do not allow the spot to air-dry during sample application. 3. Incubate in a humid chamber for 30 minutes with shaking (on MicroMix shaker setting 20/4). 4. Wash each spot with 5 µl of binding buffer with shaking, and remove buffer. Repeat two more times. 5. Wash each spot with 5 µl of DI water. Repeat once. 6.
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. © 2006 Bio-Rad Laboratories, Inc.
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