ABCC1 Antibody

What is ABCC1 and why are ABCC1 antibodies important in research?

ABCC1, also known as multidrug resistance-associated protein 1 (MRP1), is a 190 kDa protein encoded by the ABCC1 gene that belongs to the ATP-binding cassette (ABC) superfamily . This protein plays a critical role in drug and xenobiotic disposition in normal cells and helps protect tissues from cytotoxic insults . ABCC1 may also be referred to as ABC29, ABCC, GS-X, or MRP in scientific literature .

ABCC1 antibodies are essential research tools because:

• They enable detection and quantification of ABCC1 protein expression across diverse tissues and cell types

• They facilitate investigation of ABCC1's role in drug resistance mechanisms

• They support studies examining ABCC1's involvement in cancer progression and immune response

• They allow evaluation of ABCC1 as a potential prognostic marker as demonstrated in pan-cancer analysis

Structurally, ABCC1 protein is approximately 171.6 kilodaltons in mass , consisting of three membrane-spanning domains (MSD 0-2) and two nucleotide-binding domains (NBD 1-2) . This complex structure requires careful antibody selection based on experimental objectives.

What applications are ABCC1 antibodies validated for in laboratory settings?

ABCC1 antibodies are validated for numerous applications in molecular and cellular biology research:

Most commercially available ABCC1 antibodies are validated for multiple applications, though performance may vary by clone and supplier . When selecting an antibody, researchers should verify the specific applications for which each product has been validated.

What factors determine appropriate ABCC1 antibody selection for specific experiments?

Selecting the appropriate ABCC1 antibody requires consideration of several experimental factors:

• Species reactivity: Confirm the antibody recognizes ABCC1 in your experimental organism (human, mouse, rat, etc.)

• Application compatibility: Verify validation for your specific technique (WB, IHC, ELISA, etc.)

• Antibody type:

  • Monoclonal: Higher specificity for a single epitope

  • Polyclonal: Often provides stronger signal by recognizing multiple epitopes

• Conjugation options: Consider whether unconjugated antibodies or those with tags (FITC, Alexa Fluor, HRP, biotin) are more suitable

• Epitope location: For transmembrane proteins like ABCC1, epitope accessibility is crucial, especially when distinguishing between internal and external domains

• Validation data: Review experimental validation in your specific cell type or tissue

The search results indicate over 600 ABCC1 antibody products are available across different suppliers, with varying specifications and applications .

What controls should be included when using ABCC1 antibodies in experimental protocols?

To ensure reliable and interpretable results when using ABCC1 antibodies, incorporate these controls:

For quantitative applications such as ELISA, standard curves using recombinant ABCC1 protein should be established. When performing immunohistochemistry, include tissues known to express varying levels of ABCC1 to calibrate interpretation of staining intensity.

How can researchers validate the specificity of ABCC1 antibodies?

Validating ABCC1 antibody specificity requires a multi-faceted approach:

• Genetic validation:

  • Compare antibody signal in wild-type versus ABCC1 knockout/knockdown models

  • Assess signal in cells with CRISPR-engineered ABCC1 mutations

• Biochemical validation:

  • Peptide competition assays: Pre-incubating antibody with immunizing peptide should abolish specific binding

  • Western blot analysis: Confirm single band of expected molecular weight (approximately 190 kDa)

• Cross-reactivity testing:

  • Test against related ABC transporters (ABCC2, ABCC3, etc.)

  • Evaluate performance across different species if working with non-human models

• Multi-method confirmation:

  • Compare results from different detection methods (e.g., IF vs. WB)

  • Use multiple antibodies targeting different ABCC1 epitopes

• Bioinformatic correlation:

  • Compare antibody staining patterns with ABCC1 mRNA expression data from databases like TCGA and GTEx

How can ABCC1 antibodies be employed to study drug resistance mechanisms in cancer?

ABCC1 antibodies provide valuable tools for investigating multidrug resistance mechanisms through several methodological approaches:

• Expression-phenotype correlation studies:

  • Quantify ABCC1 levels in patient samples using IHC or Western blot

  • Correlate expression with treatment response and clinical outcomes

  • Studies have demonstrated ABCC1 involvement in resistance to various anticancer drugs in solid tumors including non-small cell lung cancer, prostate cancer, and breast cancer

• Functional transport assays:

  • Use antibodies to quantify ABCC1 expression alongside fluorescent substrate efflux assays

  • Measure the impact of ABCC1 inhibitors on drug accumulation

  • Glutathione significantly broadens the range of drugs transported by ABCC1, making this an important parameter to measure

• Mechanistic investigations:

  • Investigate co-localization with other resistance proteins

  • Examine conformation changes during drug transport

  • Study post-translational modifications affecting transport activity

• Therapeutic targeting:

  • Evaluate antibody-drug conjugates targeting ABCC1

  • Assess antibody-mediated inhibition of transport function

  • Monitor ABCC1 expression changes during treatment

ABCC1 can recognize and expel many hydrophobic and hydrophilic antineoplastic agents, leading to reduced drug accumulation and cellular resistance . Research has shown differential resistance to different anticancer drugs, highlighting the importance of comprehensive ABCC1 characterization in specific cancer contexts.

What methodologies are most effective for analyzing ABCC1 expression in relation to immune cell infiltration?

Recent research has revealed intricate associations between ABCC1 expression and immune response in cancer. Effective methodologies for investigating these relationships include:

• Single-cell RNA sequencing (scRNA-seq):

  • Enables correlation between ABCC1 expression and specific immune cell populations

  • Recent scRNA-seq analysis revealed positive correlation between ABCC1 expression in hepatocellular carcinoma cells and macrophage infiltration

• Computational immune profiling:

  • Utilizing tools such as the "estimate" R package to assess stromal and immune scores

  • Correlating ABCC1 expression with immune checkpoint genes using R packages like "reshape2," "ggpubr," and "corrplot"

• Multiplexed imaging techniques:

  • Immunohistochemical double staining: Combining ABCC1 antibodies with immune cell markers

  • Multiplex immunofluorescence for simultaneous detection of ABCC1 and multiple immune cell populations

• Flow cytometry analysis:

  • Multi-parameter analysis of ABCC1 expression alongside immune cell markers

  • Sorting of cell populations for downstream functional assays

• In vitro models:

  • Co-culture systems combining ABCC1-expressing tumor cells with immune cells

  • Using ABCC1 antibodies to track expression changes during immune cell interaction

The pan-cancer analysis demonstrated that ABCC1 expression exhibits significant associations with diverse immune-related genes and correlates with immune scores across multiple tumor types , positioning ABCC1 as a potential immunological biomarker.

How can researchers troubleshoot contradictory results when using ABCC1 antibodies?

When faced with contradictory results using ABCC1 antibodies across different experimental systems, consider these troubleshooting approaches:

• Antibody-specific factors:

  • Validate epitope accessibility in your specific experimental conditions

  • Consider antibody lot-to-lot variation

  • Test multiple antibody clones targeting different ABCC1 domains

• Sample preparation considerations:

  • Optimize protein extraction methods for membrane proteins

  • Evaluate different fixation protocols for preserved tissue

  • Adjust antigen retrieval methods for different tissue types

• Biological variables:

  • Assess whether different experimental models express different ABCC1 splice variants

  • Investigate post-translational modifications affecting epitope recognition

  • Consider the impact of tumor heterogeneity on sampling

• Technical validation:

  • Implement orthogonal detection methods (e.g., mass spectrometry)

  • Correlate protein detection with mRNA expression analysis

  • Perform functional assays to confirm biological relevance

• Experimental design factors:

  • Control for cell confluence and culture conditions

  • Account for treatment timing and duration

  • Consider microenvironmental factors affecting ABCC1 expression

The pan-cancer analysis showed varied ABCC1 expression across different cancer types , underscoring the importance of context-specific validation when investigating ABCC1 in different experimental systems.

What are the latest approaches for studying ABCC1's role in tumor microenvironment?

Cutting-edge approaches for investigating ABCC1's role in the tumor microenvironment include:

• Spatial transcriptomics with protein detection:

  • Correlating ABCC1 protein expression with spatial gene expression patterns

  • Mapping ABCC1-expressing cells relative to other cell types in the tumor microenvironment

• Advanced imaging techniques:

  • Multiplexed immunofluorescence for simultaneous detection of ABCC1 and multiple TME markers

  • Mass cytometry (CyTOF) using metal-conjugated antibodies for high-dimensional analysis

  • Imaging mass cytometry for spatial resolution at single-cell level

• 3D model systems:

  • Organoid cultures incorporating ABCC1-expressing tumor cells and stromal components

  • Patient-derived models maintaining original tumor architecture and heterogeneity

• In vivo approaches:

  • Intravital microscopy with fluorescently labeled ABCC1 antibodies

  • Serial sampling of tumors during treatment to track ABCC1 dynamics

• Extracellular vesicle analysis:

  • Studying ABCC1's presence on cancer-derived exosomes

  • Investigating how ABCC1-containing vesicles influence recipient cells in the microenvironment

Recent research has demonstrated that ABCC1 expression correlates with macrophage infiltration and various in vitro and in vivo experiments have substantiated the oncogenic role of ABCC1 in hepatocellular carcinoma , suggesting a potential role in modulating immune responses within the tumor microenvironment.

How can single-cell analysis techniques be combined with ABCC1 antibodies?

Single-cell technologies offer powerful approaches to study ABCC1-related heterogeneity in complex tissues:

• Cellular indexing of transcriptomes and epitopes (CITE-seq):

  • Simultaneously profiling ABCC1 at both mRNA and protein levels in individual cells

  • Correlating ABCC1 expression with comprehensive transcriptional signatures

• Advanced flow cytometry approaches:

  • High-dimensional flow cytometry for multi-parameter analysis of ABCC1 alongside other markers

  • Index sorting for linking single-cell phenotype with downstream molecular analysis

• Imaging-based single-cell analysis:

  • Imaging mass cytometry using metal-tagged ABCC1 antibodies

  • Highly multiplexed immunofluorescence for spatial single-cell analysis

• Functional single-cell assays:

  • Single-cell drug efflux assays combined with ABCC1 antibody labeling

  • Correlating ABCC1 expression with functional phenotypes at single-cell resolution

• Computational integration:

  • Integrating single-cell ABCC1 protein data with transcriptomic and genomic information

  • Trajectory analysis to study ABCC1 expression changes during cellular differentiation or treatment response

The search results mention scRNA-seq analysis revealing positive correlation between ABCC1 expression and macrophage infiltration in hepatocellular carcinoma , highlighting the value of single-cell approaches in understanding ABCC1's role in complex cellular ecosystems.

How can researchers integrate ABCC1 antibody data with genomic and clinical information?

Integrating ABCC1 antibody-based studies with genomic and clinical data enables more comprehensive translational research:

• Multi-omic correlation analyses:

  • Compare ABCC1 protein expression patterns with:

    • Gene expression profiles from TCGA/GTEx databases

    • Mutation and copy number alteration data from cBioPortal

    • Microsatellite instability (MSI) and tumor mutation burden (TMB) data

• Clinicopathological correlations:

  • Analyze associations between ABCC1 expression and:

    • Patient survival outcomes

    • Treatment response metrics

    • Disease progression parameters

• Biomarker development pipeline:

  • Standardize ABCC1 antibody-based assays for clinical application

  • Establish scoring systems for ABCC1 expression in patient samples

  • Validate prognostic or predictive significance in prospective studies

• Machine learning approaches:

  • Develop predictive models incorporating ABCC1 expression with other molecular features

  • Identify patient subgroups likely to benefit from ABCC1-targeted interventions

ABCC1 overexpression consistently predicts unfavorable outcomes based on TCGA data analysis , highlighting its potential value as a prognostic biomarker.

What experimental designs best evaluate ABCC1's functional impact in cancer models?

To rigorously assess ABCC1's functional impact in cancer, researchers should consider these experimental approaches:

• Genetic manipulation models:

  • CRISPR/Cas9 knockout or knockdown of ABCC1

  • Overexpression systems to model increased ABCC1 activity

  • Site-directed mutagenesis to study specific functional domains

• Pharmacological intervention studies:

  • Testing ABCC1 inhibitors in combination with chemotherapeutics

  • Evaluating drug efflux in the presence of glutathione modulators

  • Dose-response experiments with ABCC1 substrates

• Translational model systems:

  • Patient-derived xenografts with varying ABCC1 expression

  • Genetically engineered mouse models

  • 3D organoid cultures recapitulating tumor heterogeneity

• Immune interaction studies:

  • Co-culture experiments with immune cells and ABCC1-modified cancer cells

  • In vivo models evaluating immune infiltration in tumors with altered ABCC1 expression

  • Antibody-based imaging to track immune cell recruitment

The search results indicate that various in vitro and in vivo experiments have substantiated the oncogenic role of ABCC1 in hepatocellular carcinoma, along with increased macrophage recruitment , suggesting comprehensive approaches are needed to understand ABCC1's multifaceted functions in cancer.

How should researchers optimize ABCC1 antibodies for challenging applications?

Optimizing ABCC1 antibody protocols for challenging applications requires systematic method development:

• For low expression detection:

  • Signal amplification techniques (tyramide signal amplification, polymer detection systems)

  • Enhanced extraction protocols for membrane proteins

  • Longer primary antibody incubation at optimized temperatures

• For specific cellular compartments:

  • Optimized permeabilization protocols for accessing intracellular domains

  • Subcellular fractionation prior to analysis

  • Co-staining with organelle markers for precise localization

• For multiplexed detection:

  • Sequential antibody labeling and stripping protocols

  • Spectral unmixing for fluorophore separation

  • Careful antibody pairing to avoid cross-reactivity

• For quantitative analysis:

  • Calibration with recombinant protein standards

  • Digital image analysis with appropriate controls

  • Standardized protocols to minimize batch effects

• For fixed tissue analysis:

  • Optimization of fixation time and conditions

  • Antigen retrieval method comparison (heat-induced vs. enzymatic)

  • Testing multiple antibody clones on the same tissue

Systematic optimization should include titration of antibody concentrations, evaluation of different detection systems, and validation across multiple biological replicates to ensure reproducibility.

What are the emerging techniques for studying ABCC1 regulation?

Several cutting-edge techniques are enhancing our understanding of ABCC1 regulation:

• Proximity-based interaction assays:

  • Proximity ligation assay (PLA) for detecting protein-protein interactions involving ABCC1

  • BioID or APEX2 proximity labeling to identify ABCC1 interactors

  • FRET-based approaches to study ABCC1 conformational changes

• Epigenetic and transcriptional regulation:

  • ChIP-seq for identifying transcription factors regulating ABCC1

  • ATAC-seq to assess chromatin accessibility at the ABCC1 locus

  • CUT&RUN for precise mapping of protein-DNA interactions

• Post-translational modification analysis:

  • Phospho-specific antibodies to track ABCC1 activation

  • Mass spectrometry to identify novel modifications

  • Site-directed mutagenesis to evaluate functional impact of modifications

• Dynamic expression analysis:

  • Live-cell reporters for real-time ABCC1 expression tracking

  • Optogenetic control of ABCC1 expression

  • Cellular stress response monitoring in relation to ABCC1 levels

• High-throughput screening approaches:

  • CRISPR screens to identify regulators of ABCC1 expression

  • Small molecule library screening for ABCC1 modulators

  • Synthetic lethality screens in ABCC1-overexpressing cells

These techniques can provide deeper insights into the complex regulatory mechanisms controlling ABCC1 expression and function in normal and disease states.

How can ABCC1 antibodies contribute to personalized cancer treatment strategies?

ABCC1 antibodies have significant potential in advancing personalized cancer medicine through several approaches:

• Predictive biomarker development:

  • Standardized IHC assays to predict response to specific chemotherapeutics

  • Quantitative scoring systems correlating ABCC1 levels with treatment outcomes

  • Multiplex assays combining ABCC1 with other resistance markers

• Treatment resistance monitoring:

  • Serial sampling during treatment to track ABCC1 expression changes

  • Liquid biopsy approaches using circulating tumor cells

  • Correlation with drug levels and clinical response

• Therapeutic targeting:

  • Development of ABCC1-targeting antibody-drug conjugates

  • Evaluation of ABCC1 inhibitors as chemosensitizing agents

  • Immunotherapeutic approaches targeting ABCC1-expressing cells

• Patient stratification strategies:

  • Identifying high ABCC1 expressors for alternative treatment approaches

  • Combination strategies specifically for ABCC1-mediated resistance

  • Clinical trial enrichment based on ABCC1 status

The pan-cancer analysis revealed that ABCC1 overexpression consistently predicted unfavorable outcomes based on TCGA data , highlighting its potential value in patient stratification and treatment planning.

What quality control measures ensure reliable ABCC1 antibody-based assays in clinical research?

Implementing rigorous quality control is essential for translating ABCC1 antibody-based assays to clinical applications:

• Analytical validation requirements:

  • Sensitivity: Limit of detection determination

  • Specificity: Cross-reactivity testing with related proteins

  • Precision: Intra- and inter-assay coefficient of variation assessment

  • Accuracy: Recovery experiments with spiked samples

  • Linearity: Verification across concentration ranges

• Standardization protocols:

  • Reference standards inclusion

  • Positive and negative control tissues

  • Internal calibration controls

  • Lot-to-lot verification procedures

• Pre-analytical considerations:

  • Sample collection and processing standardization

  • Fixation time and conditions documentation

  • Storage duration effects assessment

  • Transport conditions monitoring

• Assay validation across laboratories:

  • Ring trials with multiple testing sites

  • External quality assessment participation

  • Standard operating procedures implementation

  • Regular proficiency testing

• Digital pathology considerations:

  • Algorithm validation for quantitative analysis

  • Scanner calibration and performance verification

  • Color calibration and standardization

  • Data storage and security protocols

Implementation of these quality control measures ensures that ABCC1 antibody-based assays provide consistent, reliable results that can be meaningfully interpreted in clinical research settings.