Ciphergen Rm: NSC 516 Tel: (404) 413-5379 Ciphergen Request Form |
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Charges : |
Basic Services | |
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Setup :
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$10.00 ($15.00) includes generation of a chip reading protocol (a set of two protocols for Low and High mass range) |
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Calibration : |
Molecular Standard Calibration - $60.00/experiment. |
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Data Analysis : |
analysis/consultation - $50.00/hour |
Charges : |
Full Service | |
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Chemical Surface. Normal phase. Whole chip: $120.00 |
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Chemical Surface. Ion-Exchange phase. Whole chip: $120.00 |
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Chemical Surface. Reverse phase. whole chip: $120.00 |
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Chemical Surface. Reactive surface. Whole chip: $150.00 |
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Biochemical Surface. Antibody-Chip. Whole chip: $230.00 |
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Calibration : |
Molecular Standard Calibration: $50.00/experiment |
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Data Analysis : |
Analysis/consultation: $50/hour |
| Time of Flight Mass Spectrophotometor based on SELDI technology. The system utilizes the biochemicophysical property of conventional solid-phase chromatography and MALDI-TOF mass-spectrophotometor to have more selective profiling of molecules. This selectivity of chips have clear advantages in profiling of proteins from complicated and unknown samples such as serum and cell extracts. • Accurate determination of molecular mass • Profiling of proteins from various cell extract and body fluids • Discovery and Identification of biomarker(s) • Micro-scale protein purification and purification protocol development for large scale purifications Websites: Ciphergen Protein Chip Reader SELDI Application Several Reviews of SELDI-TOF-MS technology |
| Steps for a SELDI Experiment Step one: choosing an array ProteinChip Arrays are available with different chromatographic properties, including hydrophobic, hydrophilic, anion exchange, cation exchange, and immobilized-metal affinity surfaces. Other ProteinChip Arrays with pre-activated surfaces are available for covalently coupling protein, DNA, RNA or other “bait” molecules by the user. Step two: sample application Crude biological samples such as serum,cell lysates, or other protein reparations, including those with high salt or detergent concentrations, can be applied directly to the ProteinChip Arrays. Application can be done manually by pipette, or by employing Ciphergen's customized configuration of the Beckman Coulter Biomek® 2000 laboratory automation station. The arrays are formatted with robot-friendly spot spacing and a “bioprocessor” rack of 12 arrays forming a standard microplate footprint. Step three: removal of unbound components After a short incubation period, unbound proteins are washed off the surface of the ProteinChip Array. Only proteins interacting with the chemistry of the array surface are retained for analysis. After washing, energy absorbing molecules are applied to the array as a final step. Step four: analysis in the ProteinChip Reader ProteinChip Arrays are analyzed in the ProteinChip Reader, a time-of-flight mass spectrometer. The mass values and signal intensities for the detected proteins and peptides can be viewed in several formats and then transferred to Ciphergen's powerful software suites for further in-depth analysis. SELDI Applications SELDI-MS is a very powerful and versatile technique which can answer many biological questions. Here is a list of few applications: Peptide mapping and Protein ID Biomaker Discovery and Validation Microscale Protein Purification and Purification protocol developing Molecular Recognition Protein-Protein interaction Antibody-Protein interaction DNA/RNA-Protein interaction Protein Modification Assay Phosphorylation Glycosylation SELDI Sample Prep
Protein Concentration: >0.5 mg/ml (5 ug/spot). Read the Product guidelines for each specific array type to aid in choosing buffers and wash conditions. Avoid Ionic detergents such as SDS. Use non-ionic detergents, such as Triton X-100, NP40, n-Octyl-D-glucopyranoside (OGP), Tween 20, or dodecyl maltoside may be present in final concentrations up to 1%. Salt Concentration: some arrays (such as Q10, SAX2, SM10 and WCX2) are very sensitive to high salt concentrations, while other arrays (such as H4 and H5O) actually need salt to increase binding. Avoid PEG, glycerol, DEPC, DTT. Standard Buffers Binding buffers/Washing buffers – Low and High Stringency. Chip type: Q10 Low Stringent Buffer: 50 mM Tris-HCl (pH 9.0) High Stringent Buffer: 100 mM Na Acetate (pH 6.0) CM10 Low Stringent Buffer: 100 mM Na Acetate (pH 4.0) High Stringent Buffer: 50 mM HEPES (pH 7.0) IMAC30 Low Stringent Buffer: 100 mM Na phosphate (pH 7.0), 0.5 M NaCl or PBS (pH 7.2) High Stringent Buffer: same H5O Low Stringent Buffer: 10% CAN, 0.1 % TFA or 0-50% MeOH w/ or w/o 0.1 – 1% TFA High Stringent Buffer: same PS10, PS20, RS100, Ab Coupling Buffer: PBS or Na Bicarbonate (pH 8.0) Blocking Buffer: 0.5 M Ethanolamine (pH 8.0), Tris-HCl or Glycine (pH 8.0) Binding Buffer: PBS (pH 7.2) Washing Buffer: same as binding or a little stringent This information is given as a guide to the facilities and instrumentation available in the DNA/Protein Core facility at Georgia State University. If you have any concerns or thoughts about the content of this website please contact: John Houghton (404) 413-5390
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