KIR3DL2 Antibody, Biotin conjugated

What is KIR3DL2 and what is its relevance in immune function research?

KIR3DL2 (CD158k) is a killer immunoglobulin-like receptor characterized by three Ig-like domains (3D) in its extracellular domain and a long (L) cytoplasmic tail containing two immunoreceptor tyrosine-based inhibitory motifs (ITIM domains). It functions as a receptor on natural killer (NK) cells for specific HLA-A alleles, particularly A3 and A11 . KIR3DL2 plays a crucial inhibitory role by suppressing NK cell activity, thereby preventing cell lysis when engaged with its ligands .

Unlike most other KIR family members, KIR3DL2 gene transcripts are expressed by all individuals, making it a universally relevant research target . The protein exists on the cell surface as a disulfide-linked homodimer with 70 kDa, 434 amino acid subunits . From an evolutionary perspective, human KIR3DL2's extracellular domain shows 88-92% amino acid identity to KIR3DL2 of other primates, though no direct structural orthologs exist in non-primates (with mouse Ly-49 proteins serving as functional orthologs) .

To study this receptor effectively, researchers must employ specific methodologies tailored to the unique properties of KIR3DL2 and select appropriate antibody conjugates based on experimental requirements.

What are the fundamental methodological considerations when selecting KIR3DL2 antibodies for research?

When selecting KIR3DL2 antibodies for research applications, several methodological factors must be considered:

• Specificity validation: Confirm the antibody specifically detects human KIR3DL2 in direct ELISAs and does not cross-react with other KIR family members, particularly KIR3DL1 which shares 86% amino acid identity with KIR3DL2 within the extracellular domain . The clone selection is critical - for instance, clone #539304 has been validated for detecting KIR3DL2 in human samples .

• Immunogen consideration: Verify the immunogen used to generate the antibody. High-quality antibodies are often generated using properly folded proteins, such as those from BaF3 mouse pro-B cell lines transfected with human KIR3DL2/CD158k (Accession # P43630) .

• Application compatibility: Different experimental techniques require specific antibody properties:

  • For flow cytometry: Fluorophore-conjugated antibodies provide direct detection

  • For ELISA: Biotin-conjugated antibodies offer signal amplification capabilities

  • For multiplex approaches: Select antibodies with compatible spectral properties

• Clone characteristics: Consider the antibody isotype (typically Mouse IgG for many commercial KIR3DL2 antibodies) and clonality (monoclonal antibodies provide more consistent results across experiments) .

Methodologically, researchers should validate any new antibody in their specific experimental system before proceeding with larger studies, particularly when working with primary human samples where KIR3DL2 expression may be heterogeneous.

How can researchers optimize flow cytometry protocols using biotin-conjugated KIR3DL2 antibodies?

Optimizing flow cytometry protocols with biotin-conjugated KIR3DL2 antibodies requires several methodological considerations:

• Two-step staining procedure: Unlike direct fluorophore conjugates, biotin-conjugated antibodies require a secondary streptavidin-fluorophore step. Implement sequential staining:

  • First incubate cells with biotin-conjugated KIR3DL2 antibody

  • Wash thoroughly to remove unbound primary antibody

  • Follow with streptavidin conjugated to an appropriate fluorophore

• Panel design considerations: When working with streptavidin-based detection, carefully consider fluorophore selection to minimize spectral overlap with other markers. For example, when analyzing NK cells, pair KIR3DL2 detection with CD56 markers but in different fluorescence channels to avoid compensation issues .

• Blocking endogenous biotin: Human peripheral blood mononuclear cells (PBMCs) contain endogenous biotin that can interfere with detection. Pretreat samples with avidin or streptavidin blocking reagents before adding biotin-conjugated antibodies.

• Titration optimization: Determine optimal antibody concentration through titration experiments. For human PBMCs, start with manufacturer-recommended dilutions and adjust based on signal-to-noise ratio for your specific application .

• Gating strategy example: For detecting KIR3DL2+ cells within human PBMCs, implement a stepwise gating approach:

  • Gate on lymphocytes based on FSC/SSC properties

  • Exclude doublets and dead cells

  • Gate on CD3+ T cells or CD56+ NK cells

  • Analyze KIR3DL2 expression within these populations

When analyzing results, quadrant markers should be set based on appropriate isotype control antibodies to distinguish positive from negative populations .

What methodological approaches are effective for analyzing KIR3DL2 expression in cutaneous T-cell lymphomas?

Analysis of KIR3DL2 expression in cutaneous T-cell lymphomas (CTCLs), particularly Sézary syndrome (SS), requires specialized methodological approaches:

• Flow cytometric analysis: For blood samples from CTCL patients:

  • Use a combination of T-cell markers (CD3, CD4, CD26) with KIR3DL2 antibodies

  • KIR3DL2 expression serves as the most sensitive diagnostic criterion for SS compared to all other biological criteria

  • Quantify the percentage of KIR3DL2+ cells among CD3+ T cells, as percentages >85% correlate with reduced disease-specific survival

• Tissue immunohistochemistry: For skin biopsy samples:

  • Use KIR3DL2-specific monoclonal antibodies (such as MOG1-MK323-12B11) that do not cross-react with KIR3DL1

  • Ensure proper antigen retrieval techniques for formalin-fixed, paraffin-embedded tissues

  • Quantify the percentage of KIR3DL2+ cells within the dermal infiltrate

• Molecular detection: Supplement protein expression analysis with molecular techniques:

  • Implement reverse transcription polymerase chain reaction (RT-PCR) to detect KIR3DL2 gene expression

  • Consider multiplexed assays that combine KIR3DL2 with other diagnostic markers like PLS3, Twist, and NKp46

• Longitudinal monitoring: For patient follow-up:

  • Track KIR3DL2 expression during treatment to assess therapeutic response

  • Use KIR3DL2 analysis to detect residual disease following treatment, even when clinical remission is achieved and circulating Sézary cells are undetectable by cytomorphologic analysis

These methodological approaches have demonstrated that KIR3DL2 expression is not limited to SS but is also found in transformed mycosis fungoides (tMF) and primary cutaneous anaplastic large cell lymphoma (cALCL), suggesting broader relevance in CTCL research .

What are the best practices for validating specificity of KIR3DL2 antibodies in experimental setups?

Validating the specificity of KIR3DL2 antibodies requires a multi-faceted approach to ensure experimental results accurately reflect KIR3DL2 biology:

• Cross-reactivity testing: KIR family members share significant homology, particularly KIR3DL1 which has 86% amino acid identity with KIR3DL2 in the extracellular domain . Implement:

  • Comparative staining of cell lines expressing individual KIR family members

  • Competitive binding assays with unlabeled antibodies to demonstrate specificity

  • Testing with clone-specific antibodies known not to cross-react (e.g., MOG1-MK323-12B11 for KIR3DL2)

• Cellular validation:

  • Test antibodies on human PBMCs where KIR3DL2 is expressed on a subset of NK cells

  • Confirm expression patterns match expected distribution (e.g., co-staining with CD56 for NK cells)

  • Include paired isotype controls to establish background staining levels

• Molecular validation:

  • Correlate protein detection with mRNA expression using RT-PCR or RNA sequencing

  • Validate in KIR3DL2 knockdown or knockout systems if available

  • Consider transfected cell lines (such as BaF3 mouse pro-B cells transfected with human KIR3DL2) as positive controls

• Functional validation:

  • Test antibody blockade of KIR3DL2 interaction with known ligands (HLA-A3, HLA-A11)

  • Assess effects on NK cell functions such as cytotoxicity or cytokine production

  • Verify that observed functional effects align with KIR3DL2's known inhibitory activity

Documenting these validation steps thoroughly is essential for ensuring experimental reproducibility and meaningful data interpretation in KIR3DL2 research.

How can biotin-conjugated KIR3DL2 antibodies contribute to diagnostic strategies for Sézary syndrome?

Biotin-conjugated KIR3DL2 antibodies offer specific advantages in diagnostic strategies for Sézary syndrome (SS) through their versatility in multiple detection platforms:

• Enhanced sensitivity in multiple detection systems:

  • In flow cytometry, biotin-streptavidin amplification systems can detect low-level KIR3DL2 expression in early disease stages

  • In immunohistochemistry applications, biotin-conjugated antibodies facilitate signal enhancement for improved visualization in skin biopsies

  • In multiplex detection systems, biotin-conjugated antibodies can be combined with other diagnostic markers

• Diagnostic accuracy: KIR3DL2 has been established as the most sensitive diagnostic criterion for Sézary syndrome compared to all other available biological criteria . Specifically:

  • The percentage of KIR3DL2+ cells among CD3+ T cells provides superior diagnostic performance

  • KIR3DL2 detection allows for identification of malignant clonal cell populations

• Prognostic value:

  • Multivariate analyses have established that a percentage of KIR3DL2+ cells within CD3+ T cells >85% at diagnosis is associated with significantly reduced disease-specific survival

  • This prognostic indicator can be effectively measured using various KIR3DL2 antibody conjugates, including biotin-conjugated versions

• Methodological implementation:

  • For diagnostic applications, samples should be processed within 24 hours of collection

  • Standardized protocols for staining, washing, and detection should be established for consistent results

  • Cutoff values for KIR3DL2 positivity should be determined based on institutional validation with appropriate controls

The clinical utility of KIR3DL2 detection extends beyond initial diagnosis to treatment monitoring, where it allows assessment of treatment efficiency and specificity toward tumor cells, detection of residual disease, and early identification of relapse .

What methodological approaches facilitate monitoring treatment response using KIR3DL2 as a biomarker?

Monitoring treatment response in CTCL patients using KIR3DL2 as a biomarker requires robust methodological approaches:

• Quantitative flow cytometry protocol:

  • Establish baseline KIR3DL2 expression levels before treatment initiation

  • Implement standardized staining protocols using calibrated antibody concentrations

  • Use consistent gating strategies across time points to ensure comparability

  • Express results as both percentage of KIR3DL2+ cells among CD3+ T cells and absolute counts of KIR3DL2+ cells

• Sensitivity for residual disease detection:

  • KIR3DL2 immunostaining allows detection of residual disease following treatment, even when patients clinically experience complete remission and/or have undetectable circulating Sézary cells by conventional cytomorphologic analysis

  • This superior sensitivity enables earlier intervention for emerging relapse

• Longitudinal monitoring framework:

  • Establish a schedule for periodic assessment based on treatment regimen and risk stratification

  • During active therapy: more frequent monitoring (e.g., every 1-2 months)

  • During remission: regular surveillance (e.g., every 3-6 months)

  • Upon suspicious symptoms: immediate evaluation

• Integrated assessment approach:

  • Combine KIR3DL2 expression data with other clinical parameters including:

    • Clinical skin assessment (modified SWAT score)

    • Lymph node evaluation

    • Additional flow cytometry markers

  • Correlate changes in KIR3DL2+ cell populations with treatment-specific expected timelines for response

• Response criteria standardization:

  • Define threshold values for significant change in KIR3DL2 expression

  • Categorize responses as complete molecular response, partial molecular response, stable disease, or progressive disease based on quantitative changes

This methodological framework facilitates the assessment of treatment efficiency and specificity toward tumor cells throughout the treatment course, providing valuable information for clinical decision-making .

What are the methodological considerations for implementing KIR3DL2 detection in multi-center clinical trials?

Implementing KIR3DL2 detection methods across multiple clinical sites requires systematic standardization to ensure comparable results:

• Standardized antibody selection and validation:

  • Establish a single source and lot of biotin-conjugated KIR3DL2 antibody for all sites

  • Implement a centralized antibody validation process prior to distribution

  • Provide detailed specifications including clone number, immunogen information, and validation data

• Protocol harmonization:

  • Develop standardized operating procedures (SOPs) for sample collection, processing, staining, and analysis

  • Specify critical parameters including:

    • Anticoagulant type for blood collection

    • Maximum time from collection to processing

    • Cell isolation method

    • Antibody concentrations and incubation conditions

    • Wash procedures and buffer composition

    • Data acquisition parameters

• Quality control and proficiency testing:

  • Implement a central quality control program with:

    • Stabilized control samples distributed to all sites

    • Regular proficiency testing with unknown samples

    • External quality assessment program participation

  • Establish acceptance criteria for site participation and data inclusion

• Data standardization and centralization:

  • Define uniform gating strategies and reporting formats

  • Implement centralized review of flow cytometry files

  • Utilize standardized templates for data reporting

  • Consider implementing automated analysis algorithms to reduce operator dependence

• Site training and certification:

  • Conduct initial and refresher training sessions for all laboratory personnel

  • Require demonstration of proficiency before site activation

  • Implement regular performance audits throughout the trial duration

These methodological considerations are particularly important in clinical trials evaluating KIR3DL2-targeted therapies, such as the monoclonal antibody IPH4102, which has shown promising clinical activity in early-phase studies .

How do different epitope targeting strategies affect the performance of KIR3DL2 antibodies in various applications?

The epitope targeting strategy significantly impacts KIR3DL2 antibody performance across different research applications:

• Extracellular domain (ECD) targeting:

  • Most commercial antibodies target epitopes within the three Ig-like domains of KIR3DL2's extracellular region

  • Different ECD epitopes may be variably accessible depending on KIR3DL2's conformational state or interaction with ligands

  • Antibodies targeting epitopes involved in HLA binding (such as those recognizing residues that interact with HLA-A3 or HLA-A11) may have functional blocking properties but potentially lower detection sensitivity when the receptor is ligand-engaged

• Clone-specific performance characteristics:

  • Clone #539304 (used in several commercial antibodies) was generated using BaF3 mouse pro-B cell line transfected with human KIR3DL2/CD158k and has demonstrated reliable detection in flow cytometry applications

  • The MOG1-MK323-12B11 monoclonal antibody has high specificity for KIR3DL2 without cross-reactivity to KIR3DL1, making it valuable for immunohistochemistry applications in tissue samples

  • Some clones (like AZ158) may cross-react with KIR3DL1 due to the 86% amino acid sequence identity in the extracellular domain, potentially complicating data interpretation

• Technical implications for specific applications:

  • For flow cytometry: Epitopes must remain accessible after standard fixation procedures; membrane-proximal epitopes may provide more consistent staining

  • For ELISA: Linear versus conformational epitopes significantly impact detection sensitivity in plate-based assays

  • For immunoprecipitation: Epitopes should be accessible in native conditions and not affected by detergent treatment

• Polymorphism considerations:

  • KIR3DL2 is highly polymorphic with twelve alleles identified and up to five single amino acid polymorphisms in a single individual

  • Antibodies targeting conserved regions provide more consistent detection across diverse populations

  • For research focused on specific allelic variants, epitope selection should consider known polymorphic regions

Understanding these epitope-dependent characteristics is essential for selecting the appropriate KIR3DL2 antibody for specific research questions and technical applications.

What are the critical factors for optimizing multiplex assays that include biotin-conjugated KIR3DL2 antibodies?

Optimizing multiplex assays incorporating biotin-conjugated KIR3DL2 antibodies requires addressing several critical factors:

• Strategic panel design:

  • Consider the biotin-streptavidin detection system's fluorescence properties when designing multiplex panels

  • Place the streptavidin-fluorophore in a channel with minimal spillover from other markers

  • When studying NK cells or T cells, pair KIR3DL2 detection with lineage markers (CD56, CD3) in non-overlapping channels

  • Implement a titration matrix to determine optimal concentrations in the multiplex context

• Sequential staining protocols:

  • For flow cytometry applications, implement a sequential staining approach:

    • First stain with directly conjugated antibodies

    • Wash thoroughly

    • Apply biotin-conjugated KIR3DL2 antibody

    • Wash again

    • Add streptavidin-fluorophore conjugate

  • This prevents potential cross-binding between streptavidin and biotinylated secondary antibodies

• Blocking strategy optimization:

  • Address endogenous biotin with a biotin-blocking step prior to adding biotin-conjugated antibodies

  • Implement Fc receptor blocking to prevent non-specific binding, particularly important in samples with activated immune cells

  • Consider species-specific protein blocking to reduce background

• Compensation and spectral overlap management:

  • Prepare single-stained controls for each fluorophore, including the streptavidin-fluorophore used with biotin-KIR3DL2

  • For spectral cytometry, create a comprehensive spectral library including the streptavidin-fluorophore

  • Verify compensation matrix validity specifically for the biotin-streptavidin detection system

• Validation in complex samples:

  • Test the optimized panel on samples with known KIR3DL2 expression patterns

  • Compare results with single-stained controls to ensure detection sensitivity is not compromised

  • Assess potential interference between markers, particularly for antigens co-expressed with KIR3DL2

This methodological framework is particularly valuable for comprehensive immunophenotyping of samples from patients with Sézary syndrome, where multiple markers must be analyzed simultaneously to characterize the malignant T-cell population .

How can researchers effectively address sample-specific challenges when using KIR3DL2 antibodies in diverse human specimens?

Researchers face distinct methodological challenges when applying KIR3DL2 antibodies across different human specimen types:

• Peripheral blood samples:

  • Fresh vs. cryopreserved considerations:

    • Fresh samples typically yield optimal staining for KIR3DL2

    • If cryopreservation is necessary, validate antibody performance on frozen/thawed cells specifically

    • Allow sufficient recovery time (typically 1-2 hours) after thawing before antibody staining

  • Anticoagulant effects:

    • EDTA preservation maintains better surface antigen integrity for KIR3DL2 compared to heparin

    • Process samples within 24 hours of collection for optimal results

• Skin biopsy specimens:

  • Fresh vs. fixed tissue considerations:

    • Fresh tissue yields superior flow cytometric results

    • For fixed tissue, optimized antigen retrieval protocols are critical

    • Epitope accessibility may vary between different KIR3DL2 antibody clones in FFPE samples

  • Cell dissociation protocol optimization:

    • Enzymatic dissociation must balance cell recovery with preservation of KIR3DL2 epitopes

    • Some enzymatic cocktails may cleave or modify KIR3DL2, affecting antibody binding

• Bone marrow samples:

  • High autofluorescence management:

    • Implement dead cell exclusion dyes

    • Consider fluorophores with emission spectra distant from typical autofluorescence

  • Red blood cell lysis considerations:

    • Optimize lysis procedures to minimize damage to KIR3DL2-expressing cells

    • Validate antibody performance specifically after lysis procedures

• Solid tissue samples from non-cutaneous sources:

  • Tissue-specific fixation protocol adjustments:

    • Duration of fixation impacts epitope preservation

    • Buffer composition for antigen retrieval may need tissue-specific optimization

  • Background reduction strategies:

    • Implement tissue-specific blocking protocols

    • Consider tyramide signal amplification for low-abundance detection

• Methodological validation framework:

  • For each sample type, establish:

    • Expected positive and negative cell populations

    • Appropriate control samples

    • Sample-specific optimization protocols

    • Acceptance criteria for data quality

This systematic approach to sample-specific challenges ensures reliable KIR3DL2 detection across diverse human specimens, particularly important when studying diseases with heterogeneous tissue involvement like CTCLs .

What are the common technical issues and solutions when working with biotin-conjugated KIR3DL2 antibodies?

Researchers frequently encounter several technical challenges when working with biotin-conjugated KIR3DL2 antibodies, each requiring specific methodological solutions:

• High background signal issues:

  • Problem: Non-specific binding or endogenous biotin interference

  • Solutions:

    • Implement avidin/biotin blocking kit before antibody application

    • Optimize blocking buffer composition (consider adding 1-2% BSA and 5-10% serum)

    • Increase wash steps between primary biotin-antibody and streptavidin-detection reagent

    • For immunohistochemistry, use biotin-free detection systems when endogenous biotin is problematic

• Low signal intensity:

  • Problem: Insufficient detection of KIR3DL2-positive populations

  • Solutions:

    • Optimize antibody concentration through careful titration experiments

    • Extend incubation time for both primary antibody and streptavidin-detection reagent

    • Ensure sample processing preserves surface epitopes (minimize time between collection and staining)

    • Consider signal amplification systems for low-expression samples

• Inconsistent staining across experiments:

  • Problem: Variable results between technical replicates

  • Solutions:

    • Implement strict standardization of all protocol steps

    • Use single antibody lots for related experiments

    • Prepare master mixes for reagents to minimize pipetting variations

    • Include consistent positive control samples in each experiment

• Interfering factors in clinical samples:

  • Problem: Treatment-related effects on detection sensitivity

  • Solutions:

    • Document patient treatment history prior to sample collection

    • Validate antibody performance in samples from patients on specific therapies

    • Implement appropriate waiting periods after treatment when possible

    • Consider alternative detection approaches for heavily treated samples

• Storage and stability concerns:

  • Problem: Degradation of biotin-conjugated antibodies

  • Solutions:

    • Store concentrated antibody at 2-8°C; never freeze

    • Protect from light during storage and handling

    • Prepare working dilutions fresh before each experiment

    • Validate performance periodically with control samples

These methodological solutions help ensure reliable and reproducible results when working with biotin-conjugated KIR3DL2 antibodies across various research applications.

How can researchers validate their flow cytometry protocols for KIR3DL2 detection in specialized research contexts?

Validating flow cytometry protocols for KIR3DL2 detection requires systematic methodological approaches tailored to specific research contexts:

• Protocol validation in cutaneous T-cell lymphoma research:

  • Establish reference ranges using samples from:

    • Healthy controls (expect low KIR3DL2 expression on NK cell subsets)

    • Confirmed Sézary syndrome patients (expect high KIR3DL2 expression on malignant T cells)

    • Other CTCL subtypes for differential expression patterns

  • Validation metrics should include:

    • Sensitivity: ability to detect minimum percentage of KIR3DL2+ cells

    • Specificity: correct identification of KIR3DL2+ vs. KIR3DL2- cells

    • Reproducibility: consistent results across operators and instruments

• Specialized cell population analysis:

  • For NK cell research:

    • Implement CD56 co-staining to identify KIR3DL2+ NK cell subsets

    • Include functional markers to correlate KIR3DL2 expression with NK cell activity

    • Calculate the frequency of KIR3DL2+ cells within defined NK subpopulations

  • For T cell research:

    • Use T cell lineage markers (CD3, CD4, CD8) in combination with KIR3DL2

    • In CTCL research, establish gating strategies to separate malignant from benign T cells

    • Quantify both percentage and absolute counts of KIR3DL2+ T cells

• Analytical validation framework:

  • Precision assessment:

    • Intra-assay: minimum of 10 replicates of the same sample

    • Inter-assay: minimum of 3 independent experiments over 3 different days

    • Inter-operator: minimum of 2 different operators performing identical protocols

  • Accuracy assessment:

    • Spike-in experiments with cells of known KIR3DL2 status

    • Correlation with alternative methods (e.g., RT-PCR for KIR3DL2 mRNA)

    • Comparison with reference laboratory results when available

• Clinical validation:

  • Establish clinical cutoff values through ROC curve analysis

  • Calculate sensitivity and specificity for disease detection

  • Define reporting structures that include both analytical data and interpretive comments

These validation approaches ensure that flow cytometry protocols for KIR3DL2 detection generate reliable data that can be meaningfully interpreted in various research and clinical contexts .

What research approaches can address inconsistencies in KIR3DL2 expression data across different detection platforms?

When facing inconsistencies in KIR3DL2 expression data across different detection platforms, researchers should implement systematic comparative methodologies:

• Cross-platform comparison study design:

  • Select representative samples spanning negative, low, medium, and high KIR3DL2 expression

  • Process identical aliquots in parallel using:

    • Flow cytometry with various conjugated antibodies

    • Immunohistochemistry on fixed cell preparations

    • RT-PCR for KIR3DL2 mRNA quantification

    • Protein-based methods like Western blot or ELISA

  • Analyze correlation coefficients between methods to identify systematic differences

• Technical sources of variation analysis:

  • Epitope accessibility differences:

    • Some antibody clones may detect epitopes differently affected by fixation or processing

    • Test multiple clones targeting different KIR3DL2 regions on the same samples

  • Detection threshold variations:

    • Determine minimum detectable expression for each platform

    • Establish standardized positivity thresholds calibrated across methods

  • Sample preparation effects:

    • Systematically evaluate how different preparation methods affect each detection platform

    • Develop normalized scoring systems to account for method-specific biases

• Biological sources of variation assessment:

  • Splice variant detection:

    • Design PCR primers to detect potential KIR3DL2 splice variants

    • Correlate variant expression with antibody detection efficiency

  • Post-translational modifications:

    • Investigate if glycosylation or other modifications affect antibody binding

    • Compare native vs. denatured/reduced detection efficiency

• Harmonization strategies:

  • Develop conversion algorithms between platforms after establishing correlation patterns

  • Create standardized reporting frameworks that account for platform-specific characteristics

  • Implement reference standards that can be measured across all platforms

• Integrated multi-platform approach:

  • For critical research applications, implement multiple detection methods in parallel

  • Weight results based on established reliability metrics for each platform

  • Report comprehensive KIR3DL2 profiles rather than single-platform measurements

This systematic approach to platform inconsistency not only resolves technical discrepancies but may also reveal important biological insights about KIR3DL2 expression and regulation that would be missed by single-platform approaches .