Methods and Protocols
All methods and protocols used for studies described within this report are listed. Detailed recipes for preparation of solutions are also available and specified within each protocol.
Standard AfCS Protocols:
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B-Cell Preparation and Characterization |
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Isolation of Resting B Lymphocytes from One or More Groups of Four Mouse Spleens |
PP0000000100 |
| Isolation of Resting B Lymphocytes from Sixteen Mouse Spleens | PP0000001600 |
| Maintenance of the AutoMACS Cell Sorter | PP0000001700 |
| Characterization of Cells by Flow Cytometry | PP0000001800 |
| Live/Dead Assay for Cell Viability | PP0000002300 |
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Quantification of Apoptosis and Cell Cycle Distribution of Primary B Cells Using Propidium Iodide |
PP0000002400 |
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Microarray Analysis |
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| Preparation of B-Lymphocyte RNA for Microarray Analysis | PP0000000900 |
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Visualization of RNA Preparations on 1% Agarose Gels |
PP0000002500 |
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Functional Responses |
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Assay of Intracellular Free Calcium in Suspended B Cells (with Fluorskan Ascent) |
PP0000001100 |
| Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination | PP0000001200 |
| Assay of Cyclic AMP in Lysates of Cells | PP0000001500 |
| Preparation of B-Lymphocyte Lysates for Western Blot Analysis | PP0000001000 |
| Preparation of Positive Control A for Western Blot Analysis | PS0000008300 |
| Western Blot Analysis-Phosphoprotein-Specific Antibody Mixture | PP0000000700 |
| SDS-PAGE-Bio-Rad Criterion Cell | PP0000000300 |
| Protein Transfer-Bio-Rad Criterion Blotter | PP0000000500 |
Additional Methods or Protocols:
| Affymetrix GeneChip Expression Analysis Manual | [PDF] |