ACCS Antibody

What is ACCS protein and why are antibodies against it important for research?

ACCS, or 1-aminocyclopropane-1-carboxylate synthase-like protein 1, is a 57324 MW protein belonging to the class-I pyridoxal-phosphate-dependent aminotransferase family . Also known as ACC synthase-like protein 1, ACCS has several synonyms including PHACS . This protein plays important roles in metabolic pathways related to aminotransferase activity.

Anti-ACCS antibodies are critical research tools that enable:

• Detection of ACCS expression in various cell and tissue types

• Analysis of ACCS protein levels under different experimental conditions

• Investigation of ACCS localization within cells and tissues

• Studying potential roles of ACCS in disease mechanisms

It's important to note that there is potential confusion in nomenclature, as ACC can also refer to other proteins in the literature, including BMS1 ribosome biogenesis factor , which highlights the importance of antibody specificity validation.

What applications are ACCS antibodies most commonly used for?

ACCS antibodies are validated and optimized for multiple applications in molecular and cellular biology research:

• Western Blot (WB): Recommended dilutions typically range from 1:500-1:2000 . This technique allows detection of ACCS protein in cell or tissue lysates with high specificity.

• Immunohistochemistry (IHC): Used at dilutions of 1:50-1:200 , enabling visualization of ACCS in fixed tissue sections.

• ELISA: Allows quantitative measurement of ACCS in solution.

• Immunocytochemistry (ICC): Enables detection of ACCS in cultured cells.

• Immunofluorescence: Combines with fluorescent secondary detection systems for visualization of ACCS localization.

Each application requires specific optimization of antibody concentration, incubation conditions, and detection methods to achieve reliable results.

What species reactivity should be considered when selecting an ACCS antibody?

When selecting an ACCS antibody, species reactivity is a critical consideration:

• Many commercially available ACCS antibodies react with human, mouse, and rat ACCS proteins

• Some antibodies are specific to human ACCS only

• Cross-species reactivity varies between products and should be verified prior to use

Researchers should carefully match the antibody's species reactivity to their experimental model. For example, antibody A10077 from Boster is reactive to ACCS in human, mouse, and rat , making it versatile for comparative studies across these species.

Sequence homology between species should also be considered, as regions of high conservation may result in cross-reactivity even when not explicitly stated in product specifications.

How can I validate the specificity of an ACCS antibody for my research?

Rigorous validation is essential for reliable results with ACCS antibodies. A comprehensive validation approach includes:

• Positive controls: Use cell lines or tissues known to express ACCS

• Negative controls: Include samples lacking ACCS expression

• Peptide competition assays: Pre-incubate antibody with immunizing peptide to block specific binding

• Multiple antibody validation: Test different antibodies targeting distinct ACCS epitopes

• Knockout/knockdown verification: Test in samples with genetically reduced ACCS

Reputable manufacturers validate antibodies across multiple applications. For example, Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive and negative samples to ensure specificity and high affinity .

Additionally, bioinformatic approaches are increasingly important in antibody validation. Text mining methods can extract statements about antibody specificity issues from literature to create knowledge bases alerting users about potentially problematic antibodies .

What factors should I consider when choosing between monoclonal and polyclonal ACCS antibodies?

Selection depends on your specific research needs:

• Choose monoclonal antibodies (like Mouse Anti-ACCS Recombinant Antibody from Creative Biolabs ) when highest specificity is required

• Consider polyclonal antibodies (like Rabbit Polyclonal Anti-ACCS Antibody ) when detecting proteins with low expression levels or when epitope accessibility may be compromised

How important is the immunogen information when selecting an ACCS antibody?

Immunogen information is crucial for several reasons:

• Epitope prediction: Helps predict which region of ACCS the antibody will recognize

• Cross-reactivity assessment: Allows evaluation of potential cross-reactivity with similar proteins

• Application compatibility: Some epitopes are more suitable for certain applications (e.g., conformational epitopes for native applications)

• Blocking experiments: Essential information for designing peptide competition assays

For example, the BosterBio antibody uses recombinant full-length Human ACCS as the immunogen , suggesting it might recognize multiple epitopes throughout the protein. This comprehensive coverage can be advantageous for detection but might increase risk of cross-reactivity with related proteins.

When selecting an antibody, researchers should evaluate whether the immunogen represents the full protein or a specific peptide region, and consider how this matches their experimental needs.

What are the optimal conditions for using ACCS antibodies in Western blot?

For optimal Western blot results with ACCS antibodies, follow these methodological guidelines:

• Sample preparation:

  • Use appropriate lysis buffers that preserve protein integrity

  • Include protease inhibitors to prevent degradation

  • Denature samples completely (ACCS MW: ~57 kDa )

• Gel electrophoresis:

  • Use 10% SDS-PAGE gels suitable for ~57 kDa proteins

  • Load 20-50 μg total protein per lane

• Transfer conditions:

  • Transfer to PVDF or nitrocellulose membranes

  • Use standard transfer conditions (100V for 1 hour or 30V overnight)

• Blocking:

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

• Antibody incubation:

  • Dilute primary antibody 1:500-1:2000 in blocking buffer

  • Incubate overnight at 4°C with gentle agitation

  • Wash 3x with TBST, 5 minutes each

  • Incubate with appropriate HRP-conjugated secondary antibody (1:5000-1:10000)

  • Wash 3x with TBST, 10 minutes each

• Detection:

  • Use enhanced chemiluminescence (ECL) detection reagents

  • Optimize exposure time to avoid saturation

These conditions should be optimized for each specific antibody and experimental system.

What controls should be included in experiments using ACCS antibodies?

A robust experimental design using ACCS antibodies requires multiple controls:

For IHC applications, additional tissue-specific controls should be included, and antigen retrieval methods should be optimized based on fixation conditions.

Boster validates their antibodies with known positive control and negative samples to ensure specificity and high affinity , providing a good baseline for control selection.

How should I optimize ACCS antibody dilutions for different applications?

Systematic dilution optimization is crucial for each application:

For Western Blot:

• Start with the manufacturer's recommended range (1:500-1:2000)

• Prepare a dilution series (e.g., 1:500, 1:1000, 1:2000)

• Run identical blots with positive control samples

• Compare signal-to-noise ratio at each dilution

• Select dilution with strongest specific signal and minimal background

For IHC:

• Begin with recommended dilution range (1:50-1:200)

• Test on known positive tissues with appropriate negative controls

• Evaluate specific staining versus background

• Adjust antigen retrieval methods if necessary

• Document optimal conditions for reproducibility

For other applications:

• ELISA: Perform checkerboard titration of coating antigen and primary antibody

• ICC/IF: Test multiple fixation methods alongside antibody dilutions

• Flow cytometry: Titrate to determine optimal concentration by signal separation

Document all optimization experiments methodically to ensure reproducibility across experiments.

How can computational approaches improve ACCS antibody design and selection?

Recent advances in computational biology are revolutionizing antibody engineering, with applications for ACCS antibodies:

• Sequence-structure-function prediction:

  • Computational models can predict antibody specificity from sequence data

  • Phage display experiments provide training data for these models

  • Models can design antibodies with both specific and cross-specific properties

• Data mining applications:

  • Mining antibody repertoires can discover natural antibody clones with better developability profiles

  • Dissecting public mining clues through repertoire mining illuminates paths forward

  • The Observed Antibody Space database contains over half a billion antibody sequences for comparative analysis

• Integration with experimental data:

  • Computational models can predict variants not present in training sets

  • These predictions can guide experimental testing of novel sequences

  • Integrating computational predictions with experimental validation creates powerful iterative improvement cycles

These approaches can significantly reduce the time and resources needed for antibody development and optimization while improving specificity and reducing cross-reactivity.

What are the emerging technologies in antibody-based detection relevant to ACCS research?

Several emerging technologies have potential applications for ACCS antibody-based research:

• Antibody-Cell Conjugation (ACC):

  • A new research direction in medicine and biotechnology

  • Involves chemical coupling of antibodies to cells like NK or CIK cells

  • Enables cells to have new functions through antibody targeting

  • Could potentially be applied using ACCS antibodies for targeting specific tissues or cells

• Multiplexed detection systems:

  • Simultaneous detection of multiple proteins including ACCS

  • Provides contextual information about protein interaction networks

  • Reduces sample requirements and enables spatial correlation

• Single-molecule detection:

  • Super-resolution microscopy combined with specific antibodies

  • Enables visualization of individual ACCS molecules in cellular context

  • Provides insights into molecular clustering and protein-protein interactions

• Advanced validation technologies:

  • CRISPR-based knockout validation

  • Mass spectrometry verification of antibody targets

  • Automated high-throughput antibody characterization platforms

These technologies represent the cutting edge of antibody-based research and offer new possibilities for studying ACCS in complex biological systems.

How can I use ACCS antibodies in combination with other detection methods for comprehensive analysis?

A multi-modal approach to ACCS analysis provides more robust and informative results:

• Complementary protein detection methods:

  • Combine antibody-based detection (WB, IHC) with mass spectrometry

  • Use proximity ligation assays to study ACCS interaction partners

  • Complement protein detection with RNA analysis (qPCR, RNA-seq)

• Functional validation approaches:

  • Correlate ACCS protein levels (antibody detection) with enzymatic activity assays

  • Combine localization studies (IF with ACCS antibodies) with live-cell imaging

  • Use antibody-based pull-down assays followed by interaction partner identification

• Integrated multi-omics:

  • Correlate antibody-detected ACCS levels with transcriptomic profiles

  • Integrate proteomic and metabolomic data to understand ACCS function

  • Use systems biology approaches to contextualize ACCS in broader pathways

• Orthogonal validation strategies:

  • Validate antibody findings with CRISPR-based tagging of endogenous ACCS

  • Compare antibody-detected localization with fluorescent protein fusions

  • Use in situ hybridization to correlate protein and mRNA localization

This multi-modal approach reduces reliance on antibody specificity alone and provides a more complete understanding of ACCS biology.

What are common issues with ACCS antibodies and how can I address them?

For unresolvable issues, consider alternative methods for ACCS detection or verification, such as mass spectrometry or genetic tagging approaches.

How should I interpret contradictory results from different ACCS antibodies?

When faced with contradictory results using different ACCS antibodies:

• Evaluate antibody characteristics:

  • Compare epitope regions targeted by each antibody

  • Assess validation data for each antibody

  • Consider antibody format (monoclonal vs. polyclonal)

  • Review production methods and immunogen information

• Analyze experimental conditions:

  • Determine if application conditions favor certain epitopes

  • Assess whether denaturation might affect epitope accessibility

  • Consider fixation effects on epitope recognition

• Investigate biological variables:

  • Determine if results reflect different isoforms or post-translational modifications

  • Consider cell type-specific or context-dependent expression patterns

  • Examine potential splicing variants that might affect epitope presence

• Resolution strategies:

  • Use orthogonal techniques (mass spectrometry, RNA analysis)

  • Perform genetic knockdown/knockout validation

  • Consult literature for known issues with specific antibodies

  • Design experiments to specifically address the discrepancy

Text mining methods can extract statements about antibody specificity issues from literature to help identify problematic antibodies and explain contradictory results .

How can I quantitatively analyze ACCS expression data from antibody-based experiments?

Quantitative analysis of ACCS expression requires rigorous methodological approaches:

• Western blot quantification:

  • Use digital image analysis software (ImageJ, Image Lab)

  • Normalize ACCS band intensity to loading controls

  • Include standard curves for absolute quantification

  • Ensure signal is within linear detection range

  • Perform statistical analysis across multiple biological replicates

• IHC/ICC quantification:

  • Use digital pathology software for unbiased analysis

  • Quantify staining intensity and distribution

  • Compare to validated scoring systems if available

  • Use machine learning approaches for complex pattern recognition

  • Perform blinded scoring by multiple observers

• ELISA data analysis:

  • Generate standard curves using purified ACCS protein

  • Ensure samples fall within linear range of detection

  • Calculate concentrations using regression analysis

  • Account for sample dilution factors

  • Perform appropriate statistical analysis

• Flow cytometry analysis:

  • Use appropriate gating strategies

  • Quantify using mean fluorescence intensity

  • Include fluorescence minus one (FMO) controls

  • Consider using quantitative beads for absolute quantification

For all quantitative analyses, appropriate statistical methods should be applied, and biological significance should be distinguished from statistical significance.

How might ACCS antibodies be used in combination with advanced imaging techniques?

Integration of ACCS antibodies with cutting-edge imaging approaches offers exciting research possibilities:

• Super-resolution microscopy:

  • STORM/PALM techniques with fluorophore-conjugated ACCS antibodies

  • Nanoscale resolution of ACCS localization within cellular compartments

  • Multi-color imaging to study co-localization with interaction partners

• Expansion microscopy:

  • Physical expansion of specimens labeled with ACCS antibodies

  • Enables super-resolution imaging on conventional microscopes

  • Preserves spatial relationships between ACCS and cellular structures

• Correlative light and electron microscopy (CLEM):

  • Combine fluorescence imaging of ACCS with ultrastructural context

  • Immunogold labeling for electron microscopy validation

  • Precise localization at subcellular and organellar levels

• Intravital imaging:

  • Use of compatible ACCS antibody fragments for in vivo imaging

  • Real-time visualization of ACCS dynamics in living tissues

  • Potential for therapeutic monitoring applications

• Multiplexed imaging technologies:

  • Cyclic immunofluorescence for simultaneous detection of dozens of proteins

  • Mass cytometry imaging for highly multiplexed tissue analysis

  • Spatial transcriptomics combined with protein detection

These approaches move beyond simple detection to provide spatial, temporal, and contextual information about ACCS biology.

What role might data mining play in improving ACCS antibody research?

Data mining is becoming increasingly crucial for antibody research and could significantly impact ACCS antibody applications:

• Antibody sequence optimization:

  • Mining antibody repertoires to discover natural antibody clones with improved properties

  • Contrasting naturally occurring and engineered repertoires to cement sequence-activity relationships

  • Using the Observed Antibody Space database containing over half a billion antibody sequences

• Specificity improvement:

  • Text mining literature to identify problematic antibodies and specificity issues

  • Computational prediction of cross-reactivity based on epitope sequence similarity

  • Integration with structural data to predict antigen-antibody interactions

• Application-specific optimization:

  • Mining experimental protocols to identify optimal conditions for specific applications

  • Analyzing public data repositories for ACCS expression patterns across tissues and conditions

  • Building predictive models for antibody performance in different experimental settings

• Literature-based discovery:

  • Automated extraction of ACCS-related findings from published literature

  • Identification of knowledge gaps and contradictions in existing research

  • Generation of novel hypotheses based on integrated knowledge

These data mining approaches can dramatically accelerate research progress and improve experimental design reliability.

What therapeutic and diagnostic applications might emerge from ACCS antibody research?

While current research primarily focuses on ACCS antibodies as research tools, several potential translational applications might emerge:

• Diagnostic applications:

  • Development of ACCS-based diagnostic assays if disease associations are established

  • Inclusion in multiplexed protein panels for disease classification

  • Potential biomarker for specific metabolic or cellular stress conditions

• Therapeutic targeting approaches:

  • If ACCS is validated as a disease target, therapeutic antibodies could be developed

  • Application of Antibody-Cell Conjugation (ACC) technology with ACCS antibodies

  • Targeted drug delivery using ACCS antibodies if tissue-specific expression is established

• Monitoring applications:

  • Use in pharmacodynamic studies to assess treatment effects on metabolic pathways

  • Companion diagnostic development if ACCS modulation becomes therapeutically relevant

  • Prognostic indicator development if expression correlates with disease outcomes

• Research tool evolution:

  • Development of highly specific recombinant antibody fragments

  • Creation of intrabodies for live-cell tracking of ACCS

  • Engineering of biosensor antibodies that report on ACCS activity rather than just presence

These applications represent speculative but plausible future directions for ACCS antibody research based on evolving technologies in the antibody field.

What are the key considerations for ensuring reproducible results with ACCS antibodies?

Reproducibility in ACCS antibody research requires attention to several critical factors:

• Antibody selection and validation:

  • Choose antibodies with extensive validation data

  • Independently validate specificity in your experimental system

  • Document antibody details (supplier, catalog number, lot, concentration)

• Experimental design:

  • Include all appropriate controls

  • Design experiments with sufficient statistical power

  • Blind analysis where possible to reduce bias

• Protocol standardization:

  • Document all experimental conditions in detail

  • Maintain consistent procedures across experiments

  • Use automation where possible to reduce variability

• Data reporting:

  • Report all experimental details according to field standards

  • Include raw data or make it available upon request

  • Present both positive and negative results

• Resource sharing:

  • Share detailed protocols through repositories

  • Consider antibody validation initiatives

  • Contribute to community knowledge about antibody performance

How should researchers integrate ACCS antibody data with other experimental approaches?

A comprehensive research strategy integrates ACCS antibody data with complementary approaches:

• Multi-level validation:

  • Verify protein expression (antibody detection) with mRNA expression

  • Confirm localization with orthogonal methods

  • Validate functional findings with multiple techniques

• Systems biology integration:

  • Place ACCS findings in broader pathway contexts

  • Correlate protein levels with metabolic profiles

  • Integrate with interactome data

• Translational relevance:

  • Connect basic research findings to physiological contexts

  • Consider disease relevance of ACCS expression patterns

  • Correlate with clinical parameters when appropriate

• Technological complementarity:

  • Use antibody detection alongside newer technologies

  • Apply computational approaches to interpret complex datasets

  • Develop predictive models incorporating antibody-derived data