SPBC21B10.08c Antibody

What is SPBC21D10.08c and why is it significant for research?

SPBC21D10.08c refers to a specific gene/protein in Schizosaccharomyces pombe (fission yeast), which serves as an important model organism in cell biology research. This protein is part of the extensive research into S. pombe cell wall formation and cell cycle regulation. Antibodies against this protein allow researchers to study its expression, localization, and function in various cellular processes .

The significance of studying this protein stems from the broader importance of S. pombe in understanding fundamental cellular mechanisms, including cell cycle regulation, cell wall formation, and protein degradation pathways that are conserved in higher eukaryotes, including humans.

What specifications should I look for when selecting a SPBC21D10.08c antibody?

When selecting a SPBC21D10.08c antibody, consider these critical specifications:

Most importantly, review validation data provided by the manufacturer and independent literature to confirm the antibody's performance in your intended application .

How should I validate a SPBC21D10.08c antibody before use in my experiments?

Proper antibody validation is crucial for reliable research results. Follow this step-by-step validation approach:

• Positive and negative controls:

  • Use wild-type S. pombe strains expressing SPBC21D10.08c as positive controls

  • Use deletion mutants (ΔSPBC21D10.08c) or knockdown strains as negative controls

• Multiple technique validation:

  • Perform Western blot to confirm antibody detects a protein of the expected molecular weight

  • Compare with GFP-tagged versions of the protein if available

  • Utilize immunofluorescence to confirm expected subcellular localization

• Application-specific validation:

  • For Western blots: Test different antibody dilutions to optimize signal-to-noise ratio

  • For immunoprecipitation: Verify enrichment of the target protein using mass spectrometry

  • For immunofluorescence: Compare with known localization patterns of the protein

• Documentation:

  • Record all validation data including controls, experimental conditions, and batch information

  • Include complete validation data in publications as supplementary information

Remember that antibody validation is not a one-time process but should be repeated for new antibody batches and applications .

What are the optimal protocols for using SPBC21D10.08c antibody in Western blotting with S. pombe samples?

For optimal Western blotting with SPBC21D10.08c antibody in S. pombe samples:

Sample Preparation:

• Extract total proteins using the trichloroacetic acid (TCA) method for best results with S. pombe

• For membrane proteins, consider spheroplasting of S. pombe cells first

• Use appropriate protease inhibitors to prevent degradation

• Quantify protein concentrations to ensure equal loading

Western Blot Protocol:

• Protein Separation:

  • Use 10-12% SDS-PAGE gels depending on the target protein's molecular weight

  • Load 20-30 μg of total protein per lane

  • Include molecular weight markers

• Transfer and Blocking:

  • Transfer to nitrocellulose membranes (recommended for S. pombe proteins)

  • Block with 5% non-fat milk or BSA in TBST for 1 hour at room temperature

• Antibody Incubation:

  • Dilute primary SPBC21D10.08c antibody 1:500 to 1:2000 in blocking buffer

  • Incubate overnight at 4°C with gentle shaking

  • Wash 3-5 times with TBST, 5 minutes each

  • Incubate with appropriate HRP-conjugated secondary antibody (typically anti-rabbit IgG)

  • Wash 3-5 times with TBST, 5 minutes each

• Detection:

  • Use ECL chemiluminescence system for detection

  • Image using a digital imaging system or film

  • For quantification, include appropriate loading controls (α-tubulin or Hxk2)

Critical Considerations:

• Always include positive and negative controls

• For time-course experiments, maintain consistent protocols across all time points

• Consider using recombinant tagged proteins as additional specificity controls

How can I optimize immunofluorescence protocols for SPBC21D10.08c antibody in fission yeast cells?

Optimized Immunofluorescence Protocol for S. pombe:

What considerations are important when designing co-immunoprecipitation experiments with SPBC21D10.08c antibody?

Co-Immunoprecipitation Strategy for SPBC21D10.08c:

• Experimental Design Considerations:

  • Determine if you need to preserve weak or transient interactions

  • Consider using crosslinking agents for transient interactions

  • Plan for appropriate controls (non-specific IgG, input samples, etc.)

  • Consider using tagged versions of interacting proteins for verification

• Cell Lysis Optimization:

  • Use gentle lysis buffers to preserve protein-protein interactions

    • Standard buffer: 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.5% NP-40, 1 mM EDTA

    • For membrane proteins: Consider using 1% digitonin or 0.5% CHAPS

  • Include protease inhibitors, phosphatase inhibitors, and reducing agents

  • Keep samples cold throughout the procedure

• Antibody Binding and Precipitation:

  • Pre-clear lysates with Protein A/G beads to reduce non-specific binding

  • Use 2-5 μg of SPBC21D10.08c antibody per mg of total protein

  • Incubate antibody with lysate for 2-4 hours at 4°C with gentle rotation

  • Add pre-washed Protein A or G beads (for rabbit antibodies, Protein A is often preferred)

  • Incubate overnight at 4°C with gentle rotation

• Washing and Elution:

  • Perform 4-5 washes with lysis buffer containing reduced detergent

  • Use gentle centrifugation (1000 × g for 1 minute) between washes

  • Elute proteins by boiling in SDS sample buffer or use specific peptide elution

• Analysis Methods:

  • Western blot analysis with antibodies against suspected interacting partners

  • Mass spectrometry analysis for unbiased identification of binding partners

  • Compare results from forward and reverse co-IP experiments

• Critical Controls:

  • Input sample (5-10% of starting material)

  • IgG control (non-specific antibody of same isotype)

  • Beads-only control (no antibody)

  • Reciprocal co-IP with antibodies against suspected interacting partners

• Troubleshooting Tips:

  • If no interaction is detected, consider crosslinking to stabilize transient interactions

  • Optimize salt and detergent concentrations to balance specificity and sensitivity

  • For weak interactions, consider proximity labeling approaches as alternatives

How can I use SPBC21D10.08c antibody to study proteasome and autophagy pathways in S. pombe?

The study of proteasome and autophagy pathways in S. pombe using SPBC21D10.08c antibody requires specialized approaches:

• Monitoring Protein Degradation:

  • Perform cycloheximide chase assays to measure protein half-life

    • Treat cells with cycloheximide (100 μg/ml) to inhibit protein synthesis

    • Collect samples at different time points (0, 15, 30, 60, 120 minutes)

    • Analyze SPBC21D10.08c protein levels by Western blot

  • Compare degradation rates between wild-type and proteasome mutants (e.g., mts3-1, pad1-932, pts1-732)

• Proteasome Inhibition Studies:

  • Treat cells with proteasome inhibitors (MG132 or Bortezomib)

  • Compare SPBC21D10.08c protein levels before and after treatment

  • Look for accumulation of polyubiquitinated forms of the protein

• Autophagy Pathway Analysis:

  • Study SPBC21D10.08c protein levels in autophagy-deficient strains (e.g., Δatg8)

  • Treat cells with autophagy inhibitors (e.g., 3-methyladenine) or inducers (e.g., rapamycin)

  • Add PMSF (2 mM) to inhibit vacuolar proteases for autophagy flux studies

• Advanced Microscopy Approaches:

  • Perform co-localization studies with proteasome markers (e.g., Pad1) and autophagy markers (e.g., Atg8)

  • Use live-cell imaging to track protein dynamics during normal growth and stress conditions

  • Implement super-resolution microscopy for detailed localization studies

• Stress Response Experiments:

  • Compare protein levels and localization during:

    • Nitrogen starvation (G0 phase induction)

    • Oxidative stress (H₂O₂ treatment)

    • Temperature shifts (especially important for temperature-sensitive proteasome mutants)

• Quantitative Analysis Methods:

  • Use quantitative Western blotting for protein level changes

  • Apply proteomics approaches (LC-MS/MS) for comprehensive protein degradation studies

  • Implement pulse-chase experiments to track protein synthesis and degradation rates

• Experimental Design Considerations:

  • Include specific controls for each pathway:

    • Proteasome pathway: Known proteasome substrates (e.g., Cut8)

    • Autophagy pathway: Known autophagy substrates

  • Always verify pathway involvement by genetic approaches (mutants) and chemical approaches (inhibitors)

What approaches can I use to study SPBC21D10.08c in the context of cell wall biosynthesis and septum formation?

To investigate SPBC21D10.08c in cell wall biosynthesis and septum formation:

• Cell Wall Component Analysis:

  • Analyze β-1,6-glucan content using specific antibodies or lectins

  • Measure β-1,3-glucan distribution using Calcofluor White staining

  • Quantify cell wall composition in wild-type versus SPBC21D10.08c mutants

• Septum Formation Studies:

  • Use time-lapse microscopy to monitor septum formation dynamics

  • Implement electron microscopy to examine septum ultrastructure

  • Stain with specific dyes:

    • Calcofluor White for primary septum visualization

    • Aniline blue for β-1,3-glucan detection

    • Lectin-based probes for specific glycoprotein patterns

• Genetic Interaction Analysis:

  • Construct double mutants with genes involved in:

    • β-1,6-glucan synthesis (e.g., sup11+)

    • β-1,3-glucanases (e.g., Gas family proteins)

    • β-1,3-glucan synthases (e.g., Bgs1, Bgs2, Bgs3, Bgs4)

  • Analyze synthetic lethality or rescue patterns

• Gene Expression Studies:

  • Perform transcriptome analysis during cell cycle progression

  • Focus on co-regulated genes involved in cell wall biosynthesis

  • Identify transcription factors that regulate SPBC21D10.08c expression

• Protein Localization During Cell Cycle:

  • Use the antibody for immunofluorescence at different cell cycle stages

  • Pay special attention to:

    • Localization during septum initiation

    • Redistribution during septum maturation

    • Presence during septum dissolution

• Biochemical Interaction Studies:

  • Immunoprecipitate SPBC21D10.08c and identify interacting partners

  • Focus on interactions with known cell wall biosynthesis enzymes

  • Investigate protein complexes during different cell cycle stages

• Cell Wall Integrity Pathway Analysis:

  • Examine SPBC21D10.08c regulation under cell wall stress (e.g., micafungin treatment)

  • Study protein levels after osmotic stress

  • Analyze localization changes in response to cell wall perturbations

• Advanced Method: Cell Wall Fractionation:

  • Fractionate cell walls to separate different glucan layers

  • Analyze protein distribution across these fractions

  • Determine association with specific cell wall components

How can I interpret conflicting data between antibody-based detection methods and gene expression analysis for SPBC21D10.08c?

When facing conflicts between antibody-based protein detection and gene expression data for SPBC21D10.08c, consider these analytical approaches:

• Common Causes of Discrepancies:

  Type of Discrepancy	Potential Explanations	Resolution Strategy
  High mRNA, low protein	Post-transcriptional regulation, rapid protein degradation	Measure protein half-life with cycloheximide chase experiments
  Low mRNA, high protein	Protein stability, translational efficiency, antibody cross-reactivity	Verify antibody specificity, use multiple antibodies
  Different localization patterns	Cell cycle specificity, developmental stage differences	Time-course experiments, synchronize cells
  Conflicting abundance measurements	Method sensitivity differences, antibody affinity issues	Quantify with absolute standards, use multiple techniques

• Systematic Validation Approach:

  • Verify antibody specificity using genetic controls:

    • Test in deletion or knockdown strains

    • Compare with epitope-tagged versions

  • Conduct time-course experiments to capture dynamic changes

  • Employ multiple detection methods (Western blot, immunofluorescence, flow cytometry)

  • Quantify protein using recombinant standards if possible

• Post-Transcriptional Regulation Assessment:

  • Examine microRNA regulation of SPBC21D10.08c

  • Investigate RNA-binding protein interactions

  • Analyze mRNA stability and translation efficiency

• Post-Translational Modification Analysis:

  • Identify potential modifications using phospho-specific or other PTM-specific antibodies

  • Perform proteomic analysis to identify modifications

  • Test if modifications affect antibody recognition

• Cell Cycle-Dependent Expression:

  • Synchronize cells and analyze both mRNA and protein throughout the cell cycle

  • Look for time delays between transcription and translation

  • Consider protein degradation timing

• Technical Considerations:

  • Evaluate sample preparation differences between techniques

  • Assess normalization methods used in each approach

  • Consider detection sensitivity limits for each method

• Integrated Data Analysis:

  • Combine proteomics, transcriptomics, and antibody-based detection

  • Use mathematical modeling to reconcile different data types

  • Consider biological context in interpreting discrepancies

What are the most common issues with SPBC21D10.08c antibody experiments and how can I resolve them?

Common Issues and Solutions for SPBC21D10.08c Antibody Experiments:

• No Signal or Weak Signal:

  Potential Causes:

  • Insufficient antibody concentration

  • Protein degradation during sample preparation

  • Low expression of target protein

  • Epitope masking due to protein folding or modifications

  Solutions:

  • Optimize antibody concentration (1:100 to 1:5000 dilution series)

  • Include protease inhibitors in all buffers

  • Enrich for the target protein via fractionation or immunoprecipitation

  • Try different epitope exposure methods (e.g., heat-mediated antigen retrieval)

  • Use alternative lysis methods that may better preserve the protein

• High Background:

  Potential Causes:

  • Non-specific binding of primary or secondary antibody

  • Insufficient blocking

  • Too high antibody concentration

  • Cross-reactivity with similar proteins

  Solutions:

  • Optimize blocking conditions (try BSA vs. milk, increase blocking time)

  • Increase washing steps (number and duration)

  • Reduce antibody concentration

  • Pre-absorb antibody with non-specific proteins

  • Use more stringent washing buffers (increase salt or detergent slightly)

• Multiple Bands in Western Blot:

  Potential Causes:

  • Protein degradation

  • Post-translational modifications

  • Splice variants

  • Cross-reactivity

  Solutions:

  • Use fresh samples with appropriate protease inhibitors

  • Analyze with phosphatase treatment if phosphorylation is suspected

  • Compare with recombinant protein standard

  • Perform peptide competition assay to confirm specificity

  • Test in knockout/knockdown cells to identify specific band

• Inconsistent Results Between Experiments:

  Potential Causes:

  • Antibody batch variation

  • Sample preparation inconsistencies

  • Cell cycle-dependent expression

  • Experimental condition variations

  Solutions:

  • Use the same antibody batch for related experiments when possible

  • Standardize sample preparation protocols

  • Synchronize cells for cell cycle-dependent proteins

  • Document all experimental conditions meticulously

• Poor Immunofluorescence Staining:

  Potential Causes:

  • Inadequate fixation

  • Insufficient permeabilization

  • Epitope masking

  • Antibody concentration issues

  Solutions:

  • Test different fixation methods (PFA vs. methanol)

  • Optimize permeabilization conditions

  • Try different antigen retrieval methods

  • Adjust antibody concentration and incubation times

  • Include known localization markers as controls

• Failed Immunoprecipitation:

  Potential Causes:

  • Low affinity of antibody for native protein

  • Harsh lysis conditions disrupting epitope

  • Epitope masked in protein complexes

  • Insufficient antibody amount

  Solutions:

  • Verify antibody works with native (non-denatured) protein

  • Try gentler lysis buffers

  • Cross-link protein complexes before lysis

  • Increase antibody amount or incubation time

  • Try different antibody orientation (e.g., direct coupling to beads)

How can I determine if my SPBC21D10.08c antibody batch is still effective after storage?

Evaluating Antibody Effectiveness After Storage:

• Initial Quality Assessment:

  • Check for visible precipitates or cloudiness in the antibody solution

  • Verify pH stability with pH strips if sufficient volume is available

  • Document physical appearance before proceeding

• Functional Validation Protocol:

  • Quick Western Blot Test:

    • Run a Western blot with a known positive control sample

    • Use the same protocol that previously worked well

    • Compare signal intensity and specificity to previous results

    • Include a loading control to normalize results

  • ELISA Validation (if applicable):

    • Perform a simple ELISA with known positive samples

    • Create a standard curve using serial dilutions

    • Compare sensitivity and detection range to previous data

  • Dot Blot Screening (for rapid assessment):

    • Spot 1-2 μl of positive control protein in serial dilutions

    • Proceed with standard antibody incubation and detection

    • Compare sensitivity to previous results or fresh antibody

• Sensitivity Determination:

  • Prepare serial dilutions of the antibody (e.g., 1:500, 1:1000, 1:2000, 1:5000)

  • Test each dilution under identical conditions

  • Compare optimal dilution to previously established working concentration

  • A significant shift in optimal dilution suggests degradation

• Specificity Assessment:

  • Test on both positive and negative control samples

  • Verify that the pattern of reactivity remains consistent

  • Check for emergence of new cross-reactivity or background

• Storage Recommendations for Future Use:

  • Aliquot antibodies into single-use volumes to avoid freeze-thaw cycles

  • Store at -20°C or -80°C for long-term storage

  • For short-term use (up to two weeks), 4°C storage is acceptable

  • Include proper documentation of storage conditions and test results

• Recovery Strategies for Partially Degraded Antibodies:

  • Concentrate the antibody if signal is weak but specific

  • Add stabilizing proteins (BSA) if not already present

  • Filter to remove any precipitates

  • Test higher concentrations to compensate for partial loss of activity

• Documentation and Decision Making:

  • Document all validation results systematically

  • Establish go/no-go criteria based on your specific application needs

  • Consider replacement if activity falls below 70% of original effectiveness

What criteria should I use to evaluate contradictory results between different antibody validation methods for SPBC21D10.08c?

When facing contradictory results between different antibody validation methods for SPBC21D10.08c, apply this systematic evaluation framework:

• Hierarchical Evaluation of Validation Methods:

  Validation Method	Reliability Ranking	Strengths	Limitations
  Genetic knockout/knockdown controls	Highest	Definitively identifies specific signal	Not always available for essential genes
  Orthogonal detection methods (MS, CRISPR)	Very High	Independent verification of protein identity	Requires specialized equipment
  Independent antibodies to different epitopes	High	Confirms target identity through multiple sites	May have different affinities/specificities
  Tagged protein expression	Medium-High	Direct comparison with antibody signal	Tagging may alter protein behavior
  Peptide competition	Medium	Confirms epitope specificity	Does not rule out cross-reactivity with similar epitopes
  Western blot molecular weight	Medium-Low	Basic verification of target size	Many proteins have similar molecular weights
  Manufacturer validation data	Variable	Provides baseline expectations	May not match your experimental conditions

• Application-Specific Considerations:

  • Different validation methods may be more relevant for specific applications

  • For Western blots: molecular weight, knockout controls, and peptide competition are most relevant

  • For immunofluorescence: localization pattern, knockout controls, and tagged protein comparisons are critical

  • For IP: enrichment of specific interactors and mass spectrometry verification are key

• Systematic Troubleshooting Approach:

  • Start with the most reliable validation method available for your system

  • Test multiple sample preparation methods to rule out technical artifacts

  • Investigate whether contradictions are application-specific or universal

  • Consider post-translational modifications that may affect recognition

• Decision-Making Framework:

  • When contradictions exist between methods, prioritize results from genetic controls

  • If contradictions persist, consider:

    • Using multiple antibodies and reporting all results

    • Implementing orthogonal detection methods

    • Redesigning experiments to accommodate limitations

• Reporting and Transparency:

  • Document all contradictory results thoroughly

  • Report limitations transparently in publications

  • Include complete validation data in supplementary materials

  • Specify the exact conditions under which the antibody was validated

• Advanced Resolution Strategies:

  • Epitope mapping to identify exact binding sites

  • Mass spectrometry validation of immunoprecipitated proteins

  • Cross-validation with CRISPR-engineered cell lines

  • Consultation with antibody specialists for technical input

How can I apply cutting-edge techniques like proximity labeling with SPBC21D10.08c antibody to map protein interaction networks?

Integrating SPBC21D10.08c Antibody with Proximity Labeling Techniques:

• BioID Approach:

  • Create a fusion of SPBC21D10.08c with BirA* biotin ligase

  • Express in S. pombe under native promoter or controlled conditions

  • Supplement growth media with biotin (50 μM) for 12-24 hours

  • Lyse cells under denaturing conditions to capture all interacting proteins

  • Purify biotinylated proteins using streptavidin beads

  • Identify interactors through mass spectrometry analysis

  • Validate key interactions using SPBC21D10.08c antibody by co-IP

• TurboID Implementation:

  • Generate TurboID-SPBC21D10.08c fusion (faster labeling than BioID)

  • Optimize biotin pulse length (10 minutes to 2 hours)

  • Process samples as with BioID approach

  • This method allows for temporal mapping of interactions

• APEX2 Proximity Labeling:

  • Create APEX2-SPBC21D10.08c fusion

  • Treat cells with biotin-phenol (500 μM) for 30 minutes

  • Add H₂O₂ (1 mM) for 1 minute to catalyze labeling

  • Quench immediately with antioxidants

  • This method offers superior spatial and temporal resolution

• Split-BioID for Specific Interaction Contexts:

  • Split BirA* into two fragments

  • Fuse one fragment to SPBC21D10.08c

  • Fuse the other to suspected interaction partners

  • Reconstitution of BirA* activity occurs only when proteins interact

  • This approach reduces background and increases specificity

• Validation and Analysis Strategies:

  • Create interaction network maps using bioinformatics tools

  • Classify interactors based on cellular compartments and functions

  • Perform GO term enrichment analysis

  • Compare interactome in different conditions (e.g., cell cycle stages, stress)

  • Validate key interactions with reciprocal BioID experiments

  • Confirm selected interactions with SPBC21D10.08c antibody co-IP

• Advanced Applications:

  • Combine with cell fractionation to focus on specific compartments

  • Implement with synchronized cultures to map cell cycle-dependent interactions

  • Integrate with CRISPR perturbations to identify functional dependencies

  • Combine with phosphoproteomics to link interactions with signaling events

• Technical Considerations:

  • Confirm fusion protein functionality compared to native SPBC21D10.08c

  • Optimize expression levels to minimize artifacts from overexpression

  • Include appropriate controls (untransfected, BirA* alone, unrelated protein fusions)

  • Consider inducible systems for temporal control of labeling

What strategies can I use to adapt SPBC21D10.08c antibody for super-resolution microscopy in S. pombe research?

Adapting SPBC21D10.08c Antibody for Super-Resolution Microscopy:

• Antibody Modification Strategies:

  • Direct Fluorophore Conjugation:

    • Conjugate antibody with bright, photostable fluorophores compatible with super-resolution (e.g., Alexa Fluor 647, Janelia Fluor dyes)

    • Optimize degree of labeling (DOL) to maintain binding while maximizing signal

    • Purify conjugated antibody to remove free dye

  • Click Chemistry Approach:

    • Modify antibody with clickable handles (e.g., DBCO, azide)

    • Perform bio-orthogonal reaction with complementary fluorophores

    • This allows for greater flexibility in fluorophore selection

  • Secondary Antibody Methods:

    • Use high-quality secondary antibodies specifically designed for super-resolution

    • Consider Fab fragments for reduced size and closer proximity to targets

• Specific Super-Resolution Techniques:

  • STORM/dSTORM Implementation:

    • Use buffer systems with oxygen scavengers (GLOX) and reducing agents (MEA)

    • Optimize switching buffer composition for S. pombe imaging

    • Collect 10,000-50,000 frames for reconstruction

  • PALM Adaptation:

    • Combine antibody labeling with photoactivatable/photoconvertible proteins

    • Create dual-color systems for correlative imaging

  • SIM Optimization:

    • Select high quantum yield fluorophores

    • Optimize sample mounting to minimize spherical aberration

    • Use thin sections or carefully optimized whole-cell preparations

  • Expansion Microscopy:

    • Link antibody to gel matrix before expansion

    • Validate epitope preservation after expansion

    • This technique is particularly useful for crowded structures

• S. pombe-Specific Sample Preparation:

  • Cell Wall Considerations:

    • Optimize cell wall digestion to improve antibody penetration

    • Consider partial rather than complete cell wall removal to maintain structure

    • Use nanobodies or Fab fragments for better penetration

  • Fixation Optimization:

    • Test multiple fixation protocols to preserve ultrastructure

    • Combine with cytoskeleton stabilization for structural studies

    • Validate that fixation maintains target protein localization

  • Mounting Considerations:

    • Use imaging-specific mounting media with appropriate refractive index

    • For STORM, use specifically formulated switching buffers

    • Consider hardening mounting media for long acquisitions

• Validation and Controls:

  • Compare with conventional imaging to verify localization patterns

  • Use multiple labeling approaches to confirm findings

  • Include appropriate fiducial markers for drift correction

  • Implement GFP/RFP tagged versions for correlation

• Quantitative Analysis Methods:

  • Apply cluster analysis algorithms to quantify protein organization

  • Measure co-localization at nanoscale resolution

  • Implement specialized software for super-resolution image analysis

  • Develop custom analysis pipelines for S. pombe cellular architecture

• Combining with Other Advanced Techniques:

  • Correlative light and electron microscopy (CLEM)

  • Live-cell and fixed-cell correlative imaging

  • Multi-color super-resolution for interactome mapping

  • Time-resolved super-resolution for dynamic processes

• Technical Considerations:

  • Optimize antibody concentration to achieve single-molecule density

  • Balance between specific signal and background fluorescence

  • Consider using smaller probes like nanobodies for improved resolution

  • Implement rigorous drift correction methods during image acquisition