Kingfisher Apex MACS Protocol Development for Yeast Cells
Master the Kingfisher Apex for efficient yeast cell separation: design, execute, and troubleshoot MACS protocols for diverse applications.
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Fundamentals of Kingfisher Apex and Yeast Cell MACS
Unit 1: Kingfisher Apex: The Basics
Intro to Kingfisher Apex
Magnetic Separation
Apex Components
Plates and Consumables
Software Interface
Unit 2: Yeast Cell MACS: Specifics
Yeast Cell MACS Intro
Yeast Cell Walls
Surface Markers
Non-Specific Binding
Cell Viability
Unit 3: MACS Protocol Workflow
Sample Preparation
Antibody Incubation
Magnetic Labeling
Apex Separation
Elution and Collection
Optimizing Yeast Cell Preparation for Kingfisher Apex MACS
Unit 1: Yeast Cell Wall Digestion
Enzymatic Digestion Intro
Optimizing Digestion
Mechanical Disruption
Digestion Troubleshooting
Post-Digestion Handling
Unit 2: Minimizing Non-Specific Binding
Blocking Agents
Optimizing Blocking
Buffer Optimization
Bead Handling
Controls for Binding
Unit 3: Cell Viability and Concentration
Viability Assessment
Viability Protocols
Concentration Impact
Cell Counting Methods
Adjusting Concentration
Designing and Executing MACS Protocols on the Kingfisher Apex
Unit 1: Protocol Design Fundamentals
MACS Protocol Overview
Application-Specific Design
Plate Configuration
Wash Step Optimization
Elution Strategy
Unit 2: Beads and Buffers
Magnetic Bead Selection
Buffer Selection
Blocking Strategies
Buffer Preparation
Bead Storage
Unit 3: Kingfisher Apex Execution and Troubleshooting
Programming the Apex
Instrument Setup
Monitoring Key Parameters
Troubleshooting: Low Yield
Troubleshooting: Binding
Antibody/Tag Selection, Conjugation, and Data Analysis
Unit 1: Antibody/Tag Selection for Yeast MACS
Targeting Yeast Cells
Antibody Selection
Tagging Strategies
Commercial Options
Custom Antibody Generation
Unit 2: Antibody/Tag Conjugation to Magnetic Beads
Conjugation Chemistry
Direct vs. Indirect
Optimizing Conjugation
Conjugation Validation
Troubleshooting Tips
Unit 3: Data Analysis and Interpretation
Cell Counting Methods
Viability Assays
Flow Cytometry Basics
qPCR for MACS
Microscopy Techniques