SPCC1919.04 Antibody

What is the recommended storage condition for research-grade antibodies?

Most research antibodies require careful storage to maintain their functionality. Based on the information from the search results, antibodies typically should be stored at -20 to -70°C for long-term storage (up to 12 months from date of receipt) . For short-term storage, reconstituted antibodies can be stored at 2 to 8°C under sterile conditions for approximately 1 month . It's critical to avoid repeated freeze-thaw cycles by using a manual defrost freezer, as this can significantly degrade antibody quality and performance .

How should antibody reconstitution be approached for optimal experimental results?

Proper reconstitution is essential for maintaining antibody functionality. While specific details for SPCC1919.04 antibody are not provided in the search results, general best practices include using appropriate sterile buffers (typically PBS or manufacturer-recommended buffer), reconstituting to the recommended concentration, and allowing complete dissolution without excessive agitation which might denature the antibody. After reconstitution, antibodies can typically be stored at -20 to -70°C under sterile conditions for approximately 6 months .

What applications are antibodies typically validated for in research settings?

From the search results, we can see that research antibodies are commonly validated for several applications including:

• Immunocytochemistry (ICC)

• Immunohistochemistry (IHC-P)

• Flow cytometry

• Cell culture studies

Each application requires specific validation parameters, and researchers should confirm that an antibody has been validated for their specific application before proceeding with experiments.

What controls should be included when using antibodies in immunostaining experiments?

When designing immunostaining experiments with antibodies like SPCC1919.04, researchers should include:

• Positive controls: Samples known to express the target protein

• Negative controls: Samples known not to express the target protein

• Secondary antibody-only controls: To assess non-specific binding of secondary antibodies

• Isotype controls: To evaluate potential background from the primary antibody class

The search results show examples of well-controlled experiments, such as using differentiated versus undifferentiated rat cortical stem cells as comparative samples for antibody validation , which demonstrates the importance of appropriate experimental controls.

How should antibody dilution optimization be approached for novel experimental systems?

Optimal antibody dilutions should be determined empirically for each application and experimental system. Based on the search results, manufacturers typically recommend that "optimal dilutions should be determined by each laboratory for each application" . A systematic approach involves:

• Performing a dilution series (e.g., 1:100, 1:500, 1:1000, 1:5000)

• Including all appropriate controls

• Evaluating signal-to-noise ratio at each dilution

• Selecting the dilution that provides maximum specific signal with minimal background

This optimization is particularly important when working with novel antibodies or uncharacterized experimental systems.

What methods can be used to validate antibody specificity for SPCC1919.04 protein?

Validating antibody specificity is critical for research reliability. Several approaches could be used:

• Western blot analysis comparing wild-type samples with knockout/knockdown models

• Immunoprecipitation followed by mass spectrometry

• Competitive binding assays with purified recombinant SPCC1919.04 protein

• Cross-reactivity testing against related proteins

• Comparing staining patterns with multiple antibodies against different epitopes of the same protein

The search results show examples of antibody validation through comparing expression patterns in different cell types and developmental stages .

How can researchers troubleshoot non-specific binding or high background issues with antibodies?

When experiencing high background or non-specific binding with antibodies like SPCC1919.04, researchers should systematically:

• Increase blocking duration and concentration (typically using BSA, serum, or commercial blocking solutions)

• Optimize primary antibody concentration (often lower concentrations reduce background)

• Adjust incubation conditions (temperature, duration)

• Increase wash steps frequency and duration

• Use more specific secondary antibodies

• Pre-adsorb antibodies with tissues/cells lacking the target protein

• Evaluate fixation methods that might preserve epitope accessibility while reducing non-specific binding

These methodological adjustments can significantly improve signal specificity in complex experimental systems.

What considerations are important when using antibodies for developing in vivo imaging techniques?

When adapting antibodies like SPCC1919.04 for in vivo imaging:

• Antibody format must be optimized (full IgG vs Fab fragments)

• Clearance kinetics must be evaluated

• Conjugation chemistry for imaging probes must preserve antibody functionality

• Potential immunogenicity must be assessed

• Tissue penetration capabilities must be determined

• Signal-to-background ratio in complex tissues must be optimized

The search results suggest that fluorescently labeled antibodies can be effective tools for studying cellular differentiation and function in complex biological systems .

How can multiparametric analysis be performed using antibodies against SPCC1919.04 alongside other markers?

For multiparametric analysis:

• Carefully select antibodies from different host species to avoid cross-reactivity

• Use directly conjugated primary antibodies when possible

• Implement sequential staining protocols when using antibodies from the same species

• Conduct thorough spectral compensation when using multiple fluorophores

• Include appropriate single-stain controls for each antibody

The search results demonstrate successful co-staining approaches, such as detecting Olig2 and Oligodendrocyte Marker O4 simultaneously in differentiated rat cortical stem cells using properly selected primary and secondary antibodies .

What quantitative methods are recommended for analyzing antibody-based immunofluorescence data?

For quantitative analysis of immunofluorescence data:

• Implement standardized image acquisition parameters

• Use appropriate software for automated or semi-automated analysis

• Establish clear criteria for positive signal threshold determination

• Normalize signal intensity to appropriate reference standards

• Account for potential autofluorescence through proper controls

• Consider multiple fields and biological replicates for statistical robustness

Examples from the search results show that both qualitative assessment and quantitative analysis of antibody staining can provide valuable information about protein expression patterns .

How should researchers approach contradictory results obtained with different antibody clones against the same target?

When facing contradictory results:

• Compare epitope locations for each antibody clone

• Evaluate antibody validation methods used for each clone

• Test multiple orthogonal detection methods

• Consider protein conformation, post-translational modifications, or splice variants that might affect epitope accessibility

• Assess experimental conditions that might affect epitope exposure differently between antibodies

• Consult literature for known limitations of specific antibody clones

This systematic approach helps distinguish between true biological findings and technical artifacts related to antibody performance.

How can antibodies be utilized in single-cell analysis techniques?

Antibodies can enhance single-cell analysis through:

• Integration with single-cell flow cytometry for protein expression profiling

• Combination with single-cell RNA sequencing for protein-RNA correlation

• Application in imaging mass cytometry for spatial protein mapping

• Use in proximity ligation assays for protein interaction studies at single-cell resolution

• Implementation in microfluidic platforms for antibody-based cell sorting

The search results demonstrate applications of antibodies in flow cytometry for detecting specific markers in cellular subpopulations .

What considerations are important when adapting antibodies for high-throughput screening platforms?

For high-throughput applications:

• Antibody stability under automated handling conditions must be assessed

• Reproducibility across plates and batches must be validated

• Compatibility with miniaturized assay formats must be confirmed

• Signal dynamic range must be optimized for detection systems

• Tolerance to DMSO or other compounds in screening libraries must be evaluated

• Data analysis pipelines for large-scale antibody-based screening must be established

These considerations ensure reliable results when implementing antibody-based detection in high-throughput research contexts.