ABCF2 Antibody

What is ABCF2 and why is it relevant to scientific research?

ABCF2 (ATP-binding cassette subfamily F member 2) is a protein belonging to the ABC transporter superfamily. In humans, the canonical protein has 655 amino acid residues and a mass of 72.3 kDa, with subcellular localization primarily in the mitochondria and cell membrane . ABCF2 is involved in lipid metabolism and ion transport, with particularly high expression in the placenta .

Research significance:

• Member of the GCN20 subfamily of ABC transporters

• Implicated in cancer progression and chemoresistance

• Potential biomarker for certain cancer types

• Subject to post-translational modifications including N-glycosylation and phosphorylation

What are the common applications for ABCF2 antibodies in research?

ABCF2 antibodies are versatile tools utilized across multiple experimental applications:

When selecting an application, consider the specific research question and whether qualitative or quantitative data is required .

What are the key differences between monoclonal and polyclonal ABCF2 antibodies?

When selecting between monoclonal and polyclonal ABCF2 antibodies, researchers should consider these fundamental differences:

Monoclonal ABCF2 Antibodies:

• Derived from a single B-cell clone

• Recognize a single epitope on ABCF2

• Examples include clone 2001C1 (ab50807) and OTI5D5 (TA811212)

• Advantages: High specificity, minimal batch-to-batch variation

• Limitations: May be more sensitive to epitope modifications or denaturation

Polyclonal ABCF2 Antibodies:

• Derived from multiple B-cell lineages

• Recognize multiple epitopes on ABCF2

• Examples include CAB4365 and CAB12555

• Advantages: Robust detection across various sample preparation conditions, stronger signal

• Limitations: Potential for cross-reactivity, batch-to-batch variation

The choice between monoclonal and polyclonal antibodies should be guided by the specific experimental needs, with monoclonals preferred when absolute specificity is critical, and polyclonals when signal amplification is desired .

How should I validate an ABCF2 antibody before using it in my research?

A comprehensive validation strategy for ABCF2 antibodies should include multiple approaches:

• Knockout/Knockdown Validation:

  • Use ABCF2 knockout cell lines (e.g., HEK-293T ABCF2 knockout)

  • Compare with wild-type samples to confirm antibody specificity

  • Employ siRNA knockdown to demonstrate signal reduction proportional to protein decrease

• Orthogonal Validation:

  • Compare results with independent antibodies targeting different ABCF2 epitopes

  • Confirm subcellular localization matches known patterns (mitochondria and cell membrane)

• Application-Specific Validation:

  • For Western blot: Verify band size (~71-72 kDa)

  • For IHC/IF: Include positive controls like placenta tissue or HeLa cells

  • For IP: Confirm enrichment of expected interaction partners

• Cross-Reactivity Assessment:

  • Test on non-target samples to evaluate potential cross-reactivity

  • Review antibody sequence alignment with other ABC family proteins

Validation is especially critical for ABCF2 due to its sequence similarity with other ABC transporters and the existence of up to two reported isoforms .

What are the optimal conditions for using ABCF2 antibodies in Western blotting?

Optimizing Western blot protocols for ABCF2 detection requires attention to several key parameters:

Sample Preparation:

• Use fresh lysates from cells with known ABCF2 expression (HeLa, MCF7, 293T)

• Include protease inhibitors to prevent degradation

• Expected molecular weight: 71-72 kDa

Protocol Optimization:

• Primary antibody dilution: 1:500-1:3000, depending on specific antibody

• Blocking: 5% non-fat milk or BSA in TBST

• Incubation: Overnight at 4°C typically yields best results

• Secondary antibody: Anti-species IgG HRP conjugate at 1:5000-1:10000

Positive Controls:

• HeLa cells, MCF7 cells, and HEK-293T cells consistently show ABCF2 expression

• Mouse tissues: testis and brain

Troubleshooting Tips:

• If detecting both isoforms, ensure gel separation is sufficient

• For weak signals, consider extended primary antibody incubation time

• For high background, increase washing steps and optimize blocking conditions

What methodological considerations are important when using ABCF2 antibodies for immunohistochemistry?

For successful ABCF2 immunohistochemistry staining, consider these methodological details:

Antigen Retrieval:

• Heat-induced epitope retrieval with TE buffer pH 9.0 is recommended

• Alternative: citrate buffer pH 6.0 may be used

Protocol Optimization:

• Recommended dilution range: 1:50-1:500

• Incubation time: Typically 1-2 hours at room temperature or overnight at 4°C

• Detection system: Polymer-based systems often provide optimal signal-to-noise ratio

Tissue Considerations:

• Positive control tissues: human colon cancer tissue , placenta

• Expression patterns: Cytoplasmic and sometimes membrane staining expected

• Background reduction: Endogenous peroxidase blocking and appropriate serum blocking

Validation Approaches:

• Compare staining patterns with published data

• Run parallel negative controls (omitting primary antibody)

• Consider dual staining with markers of subcellular compartments to confirm localization

How can ABCF2 antibodies be used to investigate chemoresistance in cancer?

ABCF2 has been implicated in chemoresistance mechanisms, particularly in ovarian cancer. Research approaches using ABCF2 antibodies include:

Experimental Approaches:

• Expression Correlation Studies:

  • Compare ABCF2 levels between sensitive and resistant cell lines using Western blot

  • Analyze patient samples with varying treatment responses using IHC

• Functional Studies:

  • Combine with siRNA knockdown or overexpression systems

  • Investigate cisplatin sensitivity changes following ABCF2 modulation

• Mechanistic Investigations:

  • Analyze ABCF2 expression in relation to Nrf2 activation

  • Evaluate post-translational modifications in response to treatment

Research Findings:
NRF2-overexpressing ovarian cancer cells with high ABCF2 levels show greater resistance to cisplatin-induced apoptosis compared to control cells. Conversely, NRF2 knockdown cells with reduced ABCF2 expression display increased cisplatin sensitivity . Additionally, direct ABCF2 overexpression decreases apoptosis in parental cells treated with cisplatin, while ABCF2 knockdown increases apoptosis in cisplatin-resistant cells .

This evidence suggests ABCF2 antibodies are valuable tools for investigating chemoresistance mechanisms and potentially identifying therapeutic targets to improve treatment efficacy.

What is the role of ABCF2 in heterodimeric ABC transporter formation and how can this be studied?

Recent research has identified novel heterodimeric ABC transporters involving ABCF family members. These can be investigated using specialized antibody-based approaches:

Heterodimer Detection Methods:

• Co-immunoprecipitation (Co-IP):

  • Use ABCF2 antibodies to pull down protein complexes

  • Identify interaction partners through Western blot or mass spectrometry

  • Control experiments crucial to establish specificity

• Proximity Ligation Assay (PLA):

  • Visualize protein-protein interactions in situ

  • Requires pairs of antibodies against suspected interaction partners

  • Generates fluorescent signals only when proteins are in close proximity

• NanoBRET Assays:

  • Similar to approaches used for other ABC protein heterodimer studies

  • Requires tagged constructs but can be validated with antibody approaches

While specific ABCF2 heterodimers have not been fully characterized, studies of related ABC transporters (like ABCB5β/B6 and ABCB5β/B9) provide methodological frameworks that could be applied to ABCF2 research .

How can ABCF2 antibodies be used to investigate its role as a prognostic marker in cancer?

ABCF2's potential as a prognostic marker can be systematically investigated using antibody-based approaches:

Tissue Microarray (TMA) Analysis:

• Use validated ABCF2 antibodies for IHC on cancer tissue microarrays

• Score expression as positive/negative or using intensity scales

• Correlate with clinical outcomes and survival data

Methodological Considerations:

• Standardize staining protocols and scoring systems

• Include relevant controls and blinded assessment

• Combine with other markers to develop comprehensive prognostic panels

• Consider molecular subtypes and treatment histories in analysis

Potential Applications:

• Patient stratification for clinical trials

• Treatment decision support

• Development of targeted therapies against ABCF2 or its regulatory pathways

What conjugated ABCF2 antibodies are available and when should they be used?

Various conjugated ABCF2 antibodies are available for specialized applications:

Application Guidance:

• For multicolor flow cytometry: Select conjugates based on instrument configuration and panel design

• For multiplexed imaging: Choose spectrally distinct fluorophores

• For sensitive detection: Consider biotin conjugates with streptavidin amplification systems

Commercial offerings include human ABCF2 PE-conjugated antibodies for flow cytometry and various custom conjugation options for research applications .

How can I use ABCF2 antibodies to study its subcellular localization and trafficking?

Investigating ABCF2 subcellular localization requires specialized immunostaining approaches:

Immunofluorescence Microscopy:

• Recommended antibody dilution: 1:50-1:200 for IF/ICC

• Expected localization: Mitochondria and cell membrane

• Co-staining markers:

  • Mitochondria: MitoTracker, TOMM20

  • Cell membrane: Na+/K+ ATPase, WGA

  • ER/Golgi: for trafficking studies

Advanced Imaging Approaches:

• Super-resolution microscopy (STED, STORM, PALM) for detailed subcellular distribution

• Live-cell imaging with compatible antibody formats or correlation with GFP-tagged ABCF2

• Electron microscopy immunogold labeling for ultrastructural localization

Dynamic Studies:

• Treatment-induced relocalization (e.g., stress conditions, drug exposure)

• Cell cycle-dependent changes in distribution

• Co-localization with interaction partners

Validation Controls:

• ABCF2 knockout cells for antibody specificity verification

• Multiple antibodies targeting different epitopes to confirm patterns

• Orthogonal approaches (fractionation + Western blot)

How do I interpret contradictory ABCF2 antibody staining results across different tissue types?

When facing contradictory ABCF2 staining patterns, consider these analytical approaches:

Potential Sources of Variation:

• Tissue-specific expression of isoforms:

  • Up to two different isoforms have been reported for ABCF2

  • Different antibodies may preferentially detect specific isoforms

• Differential post-translational modifications:

  • N-glycosylation and phosphorylation may affect epitope accessibility

  • Tissue-specific modifications could alter antibody binding

• Context-dependent protein interactions:

  • Protein-protein interactions may mask epitopes

  • Complex formation may differ between tissues

Resolution Strategies:

• Multiple antibody approach: Use several antibodies targeting different ABCF2 regions

• Complementary techniques: Combine IHC with Western blot of tissue lysates

• RNA verification: Correlate with RNAscope or RNA-seq data

• Mass spectrometry validation: For definitive protein identification

Case Example:
In research on endometrial and cervical cancers, ABCF2 showed different prognostic significance between cancer types. While 55.8% of cervical cancers expressed ABCF2 with prognostic relevance, 69.9% of endometrial cancers expressed ABCF2 but without prognostic correlation . This suggests tissue-specific functions of ABCF2 that should be considered when interpreting antibody staining results.

What cell lines and tissue types are most appropriate as positive controls for ABCF2 antibody validation?

For robust validation of ABCF2 antibodies, the following positive controls are recommended:

Cell Line Positive Controls:

• HeLa cells

• MCF7 cells

• HEK-293T cells

• NIH/3T3 cells

Tissue Positive Controls:

• Human placenta (high endogenous expression)

• Human colon cancer tissue

• Mouse testis and brain

Negative Controls:

• ABCF2 knockout HEK-293T cells (available commercial cell lysates)

• siRNA-treated cells showing knockdown

• Tissues with minimal ABCF2 expression (based on RNA-seq data)

When establishing positive controls in your laboratory, verify expression using multiple detection methods and consider creating standardized lysates or fixed specimens for long-term quality control.

How can I optimize ABCF2 immunoprecipitation for protein-protein interaction studies?

Optimizing ABCF2 immunoprecipitation requires attention to several key parameters:

Protocol Optimization:

• Antibody selection: Choose antibodies validated for IP applications (e.g., 10226-1-AP)

• Antibody amount: 0.5-4.0 μg for 1.0-3.0 mg of total protein lysate

• Lysis conditions: Use buffers that preserve protein-protein interactions

  • RIPA buffer may disrupt some interactions; consider NP-40 or digitonin-based buffers

  • Include phosphatase inhibitors to preserve phosphorylation-dependent interactions

Cross-linking Considerations:

• Consider reversible crosslinking for transient interactions

• DSP or formaldehyde (low concentration) can stabilize complexes

Control Experiments:

• IgG control IP to identify non-specific binding

• Reverse IP with antibodies against suspected interaction partners

• Input controls to assess IP efficiency

Case Example:
Research has demonstrated successful co-immunoprecipitation of ABC transporter heterodimers in melanoma cell lines Mel JuSo and UACC-257 using similar approaches, which could be adapted for ABCF2 studies .

What are the latest technological advances in ABCF2 antibody development and application?

Recent technological advances in antibody technology applicable to ABCF2 research include:

New Antibody Formats:

• Recombinant antibody technology ensuring batch consistency

• Single-domain antibodies (nanobodies) for accessing restricted epitopes

• Bi-specific antibodies for co-detection of ABCF2 with interaction partners

Advanced Detection Methods:

• Multiplexed imaging with cyclic immunofluorescence

• Mass cytometry (CyTOF) for high-dimensional analysis

• Proximity extension assays for sensitive protein detection

Functional Antibody Applications:

• Intrabodies for live-cell tracking of ABCF2

• Degradation-targeting chimeric antibodies for functional studies

• Antibody-based biosensors for real-time monitoring

Emerging Research Applications:

• Single-cell proteomics incorporating antibody-based detection

• Spatial transcriptomics combined with antibody staining

• AI-assisted image analysis for complex expression pattern characterization

These advances provide researchers with expanded capabilities for studying ABCF2 expression, localization, and function across various biological contexts and disease states.

How does ABCF2 expression correlate with cancer prognosis and what antibody-based methods best detect these relationships?

ABCF2's relationship with cancer prognosis varies by cancer type and can be studied using several antibody-based methods:

Cancer Type-Specific Correlations:

• Cervical Cancer: 55.8% of cases express ABCF2; positivity correlates with worse prognosis (risk ratio=1.437)

• Endometrial Cancer: 69.9% express ABCF2 but without significant prognostic correlation

• Ovarian Cancer: ABCF2 overexpression linked to cisplatin resistance

Optimal Detection Methods:

• Tissue Microarrays (TMAs): Enables high-throughput analysis across many samples

  • Recommended antibody dilution: 1:50-1:500 for IHC

  • Scoring: Consider both intensity and percentage of positive cells

• Multiplex IHC/IF: Correlate ABCF2 with other markers

  • Co-staining with proliferation markers (Ki-67)

  • Analysis with immune infiltration markers

  • Combination with treatment response indicators

• Digital Pathology Integration:

  • Quantitative image analysis for consistent scoring

  • Machine learning algorithms for pattern recognition

  • Correlation with clinical outcome data

Methodological Considerations:
The choice between cytoplasmic, membrane, or total ABCF2 scoring may influence prognostic correlations. Research indicates cytoplasmic ABCF2 expression is most commonly assessed for prognostic studies .

What is the relationship between ABCF2 and Nrf2 signaling in cancer, and how can antibodies help elucidate this pathway?

ABCF2 has been identified as an Nrf2 target gene with implications for cancer drug resistance:

Molecular Relationship:

• ABCF2 contains a functional antioxidant response element (ARE) in its promoter region

• Nrf2 activation upregulates ABCF2 expression

• This relationship contributes to cisplatin resistance in ovarian cancer cells

Antibody-Based Investigation Approaches:

• ChIP Assays:

  • Use anti-Nrf2 antibodies to immunoprecipitate chromatin

  • Detect ABCF2 promoter enrichment through PCR

  • Evaluate the functional role of the ARE

• Dual Staining:

  • Co-staining of Nrf2 and ABCF2 in tissue sections

  • Correlation of nuclear Nrf2 with ABCF2 expression levels

  • Spatial relationship in tissue context

• Pathway Analysis:

  • Western blot assessment of Nrf2-ABCF2 axis in response to oxidative stress

  • Effect of Nrf2 activators/inhibitors on ABCF2 expression

  • Correlation with other Nrf2 target genes

Research Findings:
Studies have demonstrated that NRF2-overexpressing cell lines contain high levels of ABCF2 and exhibit greater resistance to cisplatin-induced apoptosis . Conversely, NRF2 knockdown results in reduced ABCF2 levels and increased cisplatin sensitivity . Direct modulation of ABCF2 levels alone can alter cisplatin resistance, confirming its role in the Nrf2-mediated resistance pathway .

How can ABCF2 antibodies be used to study its role in non-cancer diseases and physiological processes?

While ABCF2 research has primarily focused on cancer, antibody-based approaches can explore its broader physiological roles:

Potential Physiological Functions:

• Ion transport mechanisms

• Lipid metabolism regulation

• Potential roles in drug transport or cellular detoxification

• Membrane biogenesis and homeostasis

Research Approaches:

• Developmental Biology:

  • Immunohistochemical analysis across developmental stages

  • Correlation with tissue differentiation and maturation

  • Expression in stem cells versus differentiated tissues

• Stress Response Studies:

  • ABCF2 expression modulation under various cellular stressors

  • Co-localization with stress response proteins

  • Post-translational modification changes during stress

• Physiological Models:

  • Expression in normal versus diseased tissues beyond cancer

  • Potential roles in inflammatory conditions

  • Regulation in response to hormonal signals

Disease Associations:
Though limited data exists, GeneCards indicates potential associations between ABCF2 and conditions such as Cystic Fibrosis and Osteogenesis Imperfecta, Type I , suggesting broader physiological roles that warrant investigation using antibody-based methods.

What are common problems with ABCF2 antibodies and how can they be resolved?

Researchers working with ABCF2 antibodies may encounter several technical challenges:

Common Issues and Solutions:

Mitigation Strategies:

• Enhanced Validation: Use knockout cell lines like ABCF2 knockout HEK-293T cells

• Multiple Antibody Approach: Compare results from different clones/sources

• Complementary Techniques: Verify findings with non-antibody-based methods

How should researchers evaluate and compare different commercial ABCF2 antibodies?

When selecting from various commercial ABCF2 antibodies, consider these evaluation criteria:

Key Selection Criteria:

• Validation Data Quality:

  • Look for antibodies validated in knockout systems

  • Assess the relevance of validation to your application

  • Check for orthogonal validation (e.g., multiple antibodies agreement)

• Application-Specific Performance:

  • Review data specifically for your application of interest

  • Consider antibody format requirements (e.g., unconjugated vs. conjugated)

  • Examine available dilution recommendations

• Target Specificity:

  • Epitope location and potential cross-reactivity

  • Species reactivity relevant to your research (human, mouse, rat)

  • Ability to distinguish between isoforms if relevant

• Publication Record:

  • Citation history in peer-reviewed literature

  • Use in similar experimental systems

Comparative Analysis Example:

What quality control measures should be implemented when using ABCF2 antibodies in long-term research projects?

For consistent results in extended ABCF2 research projects, implement these quality control measures:

Antibody Management:

• Aliquot antibodies upon receipt to minimize freeze-thaw cycles

• Document lot numbers and maintain records of performance

• Consider creating reference standards for batch-to-batch comparison

Routine Validation:

• Regular testing against positive and negative controls

• Periodic verification with knockout/knockdown systems

• Consistent use of standardized protocols

Comprehensive Documentation:

• Detailed record-keeping of antibody source, lot, dilution, and performance

• Imaging parameters, exposure settings, and analysis methods

• Raw data storage alongside processed results

Long-term Stability Measures:

• Storage according to manufacturer recommendations (typically -20°C)

• Monitoring for signs of degradation (reduced sensitivity, increased background)

• Consideration of more stable formats (e.g., lyophilized antibodies) for critical reagents

Inter-laboratory Standardization:

• Round-robin testing if multiple laboratories are involved

• Standard operating procedures (SOPs) for key applications

• Reference sample exchange and comparison

Implementing these quality control measures increases data reliability and facilitates meaningful comparison of results over time, particularly important for projects involving patient samples or longitudinal studies.

How might single-cell analysis technologies utilize ABCF2 antibodies for deeper insights?

Single-cell technologies offer exciting opportunities for advanced ABCF2 research:

Single-Cell Protein Analysis Applications:

• Mass Cytometry (CyTOF):

  • Metal-conjugated ABCF2 antibodies enable high-dimensional analysis

  • Correlation with dozens of other proteins simultaneously

  • Particularly useful for heterogeneous tissue analysis

• Single-Cell Western Blot:

  • Quantitative protein assessment at single-cell level

  • Reveals population heterogeneity in ABCF2 expression

  • Can detect rare subpopulations with distinct ABCF2 levels

• Imaging Mass Cytometry/CODEX:

  • Spatial distribution of ABCF2 in tissue context

  • Relationship to microenvironmental features

  • Co-expression patterns with multiple markers

Integration with Other Single-Cell Data:

• Correlation of ABCF2 protein levels with single-cell transcriptomics

• Function in specific cell states or differentiation trajectories

• Relationship to cellular metabolism at single-cell resolution

Emerging Research Questions:

• Cell-to-cell variation in ABCF2 expression within tumors

• Identification of rare ABCF2-high cells with potential stem-like properties

• Dynamics of ABCF2 expression in response to treatment at single-cell level

What new insights about ABCF2 heterodimer formation and function could antibody-based research reveal?

The discovery of heterodimeric ABC transporters opens new avenues for ABCF2 research:

Potential Research Directions:

• Identification of ABCF2 Binding Partners:

  • Co-immunoprecipitation with ABCF2-specific antibodies

  • Mass spectrometry analysis of interacting proteins

  • Validation of interactions using reciprocal co-IP

• Functional Characterization:

  • How heterodimer formation affects localization and function

  • Impact on substrate specificity and transport capacity

  • Regulation of heterodimer formation under different conditions

• Structural Studies:

  • Proximity ligation assays to confirm interactions in situ

  • Antibody epitope mapping to identify interaction domains

  • Correlation with structural predictions from homology modeling

Methodological Approaches:
Research on ABCB5β/B6 and ABCB5β/B9 heterodimers has employed NanoBRET assays and validation by immunoprecipitation in melanoma cell lines . Similar approaches could be adapted for ABCF2, potentially revealing novel heterodimeric complexes with functional significance.

Therapeutic Implications:
Understanding ABCF2 heterodimer formation could reveal new opportunities for therapeutic intervention, particularly in cancer contexts where ABCF2 contributes to drug resistance .

How might ABCF2 antibodies contribute to translational research and potential therapeutic development?

ABCF2 antibodies hold significant potential for translational applications:

Diagnostic Applications:

• Prognostic Biomarker Development:

  • Standardized IHC protocols for clinical application

  • Integration into multi-marker prognostic panels

  • Correlation with treatment response prediction

• Companion Diagnostics:

  • Identification of patients likely to benefit from specific therapies

  • Monitoring of ABCF2 expression during treatment

  • Early detection of resistance development

Therapeutic Applications:

• Antibody-Drug Conjugates (ADCs):

  • If ABCF2 is accessible at cell surface in certain contexts

  • Targeted delivery of cytotoxic agents to ABCF2-expressing cells

• Functional Inhibition:

  • Intracellular antibody delivery systems

  • Antibody fragments that disrupt ABCF2 function or interactions

  • Structure-guided development of small molecule inhibitors

Research Supporting Clinical Translation:

• Correlation of ABCF2 with clinical outcomes across cancer types

• Combination strategies targeting ABCF2 and related pathways

• Development of standardized clinical assays