LRRC3C Antibody

What is LRRC3C and what biological functions is it associated with?

LRRC3C, also known as Leucine-rich repeat-containing protein 3C, is involved in multiple biological processes including cell adhesion, protein-protein interactions, and signal transduction pathways. The protein has potential implications in various disease mechanisms, making it an attractive target for research across multiple fields including cancer biology, neurobiology, and developmental biology . The molecular weight of LRRC3C is approximately 29,314 Da . Understanding LRRC3C's functions and regulation could provide valuable insights into disease mechanisms and potentially lead to novel therapeutic approaches.

What types of LRRC3C antibodies are currently available for research?

Several types of LRRC3C antibodies are available for research purposes with different characteristics:

• Polyclonal antibodies: Generated in rabbits, these exhibit high specificity and sensitivity toward human LRRC3C. An example is the PACO63411 antibody which is purified through antigen affinity methods .

• Species-specific reactivity: Antibodies are available with reactivity against human and mouse LRRC3C proteins .

• Application-specific formulations: Different antibodies are optimized for specific applications such as Western blot (WB), ELISA, and immunohistochemistry (IHC) .

The selection of an appropriate antibody should be based on the intended experimental application and target species.

LRRC3C antibodies should be stored according to manufacturer specifications to maintain reactivity and performance. The typical storage conditions include:

• Storage buffer composition: 50% Glycerol, 0.01M PBS, pH 7.4

• Preservative: 0.03% Proclin 300

• Physical state: Liquid form

• Temperature: Most antibodies should be stored at -20°C for long-term storage

• Aliquoting: To avoid repeated freeze-thaw cycles, which can diminish antibody activity

Following these storage guidelines ensures antibody stability and consistent experimental results over time.

How should I validate the specificity of LRRC3C antibodies before experimental use?

Validating antibody specificity is crucial for obtaining reliable results. For LRRC3C antibodies, consider these validation approaches:

• Positive and negative controls: Use tissues or cell lines known to express or not express LRRC3C

• Blocking peptide experiments: Pre-incubate the antibody with the immunizing peptide (such as recombinant Human LRRC3C protein, amino acids 42-224) to confirm specific binding

• Cross-reactivity testing: Assess potential cross-reactivity with related proteins, particularly other leucine-rich repeat-containing proteins

• Multiple antibody validation: Use different antibodies targeting different epitopes of LRRC3C

• Knockout/knockdown validation: Compare staining in wild-type vs. LRRC3C-knockout or knockdown samples

The antibodies available commercially have undergone validation processes to ensure high specificity without significant cross-reactivity or interference with LRRC3C analogs .

What is the recommended protocol for using LRRC3C antibodies in immunohistochemistry?

For optimal immunohistochemical detection of LRRC3C, follow this methodological approach:

• Tissue preparation: For paraffin-embedded tissues, perform dewaxing and hydration steps

• Antigen retrieval: Use high-pressure retrieval in citrate buffer (pH 6.0) to expose epitopes

• Blocking: Block with 10% normal goat serum for 30 minutes at room temperature

• Primary antibody: Apply LRRC3C antibody at a dilution of 1:20 to 1:200 (with 1:100 being commonly used)

• Secondary antibody: Apply appropriate species-specific secondary antibody

• Detection: Use a compatible detection system (e.g., HRP-based)

• Counterstaining: Apply nuclear counterstain as needed

• Mounting: Mount slides with appropriate medium

This protocol has been successfully used with the PACO63411 antibody on human skin tissue samples using a Leica BondTM system .

What are the key considerations when performing LRRC3C ELISA assays?

When performing ELISA assays for LRRC3C quantification, consider these methodological aspects:

• Assay principle: Commercial kits utilize the sandwich enzyme immunoassay principle with pre-coated antibodies specific to LRRC3C

• Sensitivity parameters:

  • Human LRRC3C ELISA kits: 0.066 ng/mL sensitivity with detection range of 0.16-10 ng/mL

  • Mouse LRRC3C ELISA kits: 0.068 ng/mL sensitivity with detection range of 0.156-10 ng/mL

• Sample preparation: Properly process tissue homogenates and biological fluids

• Standard curve preparation: Prepare standards through serial dilution to establish a reliable standard curve

• Quality control: Include controls to assess precision (intra-assay CV%<8%, inter-assay CV%<10%)

• Incubation conditions: Follow recommended temperatures and timing for all incubation steps

• Data analysis: Compare sample OD values to the standard curve to determine LRRC3C concentrations

Following these guidelines will ensure accurate and reproducible quantification of LRRC3C in experimental samples.

How can LRRC3C antibodies be utilized to investigate protein-protein interactions?

LRRC3C contains leucine-rich repeat domains that mediate protein-protein interactions. To investigate these interactions, consider these methodological approaches:

• Co-immunoprecipitation (Co-IP):

  • Use anti-LRRC3C antibodies to pull down LRRC3C and associated proteins

  • Analyze precipitated complexes by Western blot or mass spectrometry

  • Validate interactions using reciprocal Co-IP with antibodies against suspected binding partners

• Proximity ligation assay (PLA):

  • Apply primary antibodies against LRRC3C and potential interacting proteins

  • Use species-specific PLA probes to generate signals only when proteins are in close proximity

  • Quantify interaction signals using fluorescence microscopy

• FRET/BRET analysis:

  • Create fusion proteins of LRRC3C and potential interactors with appropriate fluorophores/luciferase

  • Measure energy transfer as indication of protein proximity

• Pull-down assays:

  • Use recombinant LRRC3C proteins (available from commercial sources in E. coli expression systems)

  • Identify binding partners from cell lysates or recombinant protein libraries

These techniques provide complementary information about LRRC3C's interaction network and can help elucidate its role in signaling pathways.

What approaches can be used to study LRRC3C expression in disease models?

To investigate LRRC3C's potential role in disease mechanisms, consider these experimental approaches:

• Comparative expression analysis:

  • Use validated LRRC3C antibodies for IHC to compare expression patterns in normal versus diseased tissues

  • Quantify expression differences using Western blot with densitometry analysis

  • Measure protein levels using quantitative ELISA with sensitivity in the ng/mL range

• Functional studies:

  • Overexpress or knock down LRRC3C in disease-relevant cell models

  • Assess phenotypic changes using cellular assays

  • Monitor changes in suspected signaling pathways

• Tissue microarray analysis:

  • Screen multiple patient samples simultaneously using LRRC3C antibodies

  • Correlate expression with clinical parameters and outcomes

• Animal model studies:

  • Use species-specific antibodies (mouse LRRC3C antibodies are available)

  • Analyze temporal and spatial expression patterns during disease progression

These approaches can help establish whether LRRC3C plays a functional role in disease pathogenesis and potentially identify it as a therapeutic target.

How can I optimize LRRC3C detection in low-expression samples?

When working with samples having low LRRC3C expression, consider these optimization strategies:

• Signal amplification methods:

  • Tyramide signal amplification (TSA) for IHC applications

  • Enhanced chemiluminescence (ECL) substrates with extended duration for Western blot

  • Biotin-streptavidin systems for signal enhancement

• Sample enrichment:

  • Immunoprecipitation to concentrate LRRC3C before analysis

  • Subcellular fractionation to isolate compartments with higher LRRC3C concentration

• Detection system optimization:

  • Use high-sensitivity ELISA kits with detection limits as low as 0.066-0.068 ng/mL

  • Optimize antibody concentrations for maximum signal-to-noise ratio

  • Increase primary antibody incubation time (overnight at 4°C)

• Reducing background:

  • Optimize blocking conditions

  • Include additional washing steps

  • Use monovalent Fab fragments to block endogenous immunoglobulins in tissue samples

These techniques can significantly improve detection sensitivity while maintaining specificity for LRRC3C.

What are common issues with LRRC3C antibody experiments and how can they be resolved?

Researchers may encounter several challenges when working with LRRC3C antibodies:

When troubleshooting, make systematic changes to one variable at a time while documenting all modifications to identify the optimal conditions for LRRC3C detection.

How should I interpret and analyze LRRC3C ELISA data?

For proper interpretation of LRRC3C ELISA results, follow these analytical approaches:

• Standard curve analysis:

  • Generate a standard curve using known concentrations (typically 0.156-10 ng/mL)

  • Ensure the curve fits appropriate regression model (often 4-parameter logistic)

  • Verify R² value exceeds 0.98 for reliable quantification

• Quality control assessment:

  • Check that intra-assay CV% is <8% and inter-assay CV% is <10%

  • Verify that duplicate/triplicate measurements are consistent

  • Confirm positive controls fall within expected ranges

• Data normalization:

  • For tissue samples, normalize to total protein concentration

  • For cell culture samples, consider normalizing to cell number or housekeeping proteins

  • For complex biological fluids, evaluate whether additional normalization is needed

• Statistical analysis:

  • Apply appropriate statistical tests based on experimental design

  • Consider biological relevance of observed differences in LRRC3C levels

  • Account for multiple comparisons when performing numerous tests

This methodical approach ensures reliable quantification and meaningful biological interpretation of LRRC3C measurements.

How can I address contradictory results from different LRRC3C antibodies?

When facing contradictory results from different LRRC3C antibodies, apply this systematic approach:

• Epitope comparison:

  • Compare the immunogens used to generate each antibody

  • Determine if antibodies recognize different domains of LRRC3C (the full-length protein is 29,314 Da)

  • Consider that some antibodies target specific regions (e.g., amino acids 42-224)

• Validation strength assessment:

  • Review validation data for each antibody

  • Evaluate specificity claims and cross-reactivity testing

  • Check literature for independent validation of each antibody

• Application-specific performance:

  • Some antibodies perform better in certain applications (WB vs. IHC vs. ELISA)

  • Test antibodies at multiple dilutions for each application

  • Consider native vs. denatured protein recognition differences

• Reconciliation strategies:

  • Use orthogonal methods to validate results (mRNA expression, mass spectrometry)

  • Consider employing knockout/knockdown controls with each antibody

  • Consult with antibody manufacturers for technical support

These approaches can help determine which results are most reliable and provide a clearer understanding of LRRC3C expression or function.

What are the potential applications of LRRC3C research in disease mechanisms?

LRRC3C is implicated in several biological processes with potential relevance to disease:

• Cancer biology:

  • LRRC3C's involvement in cell adhesion suggests potential roles in tumor progression and metastasis

  • Investigation of expression patterns across cancer types could identify diagnostic or prognostic biomarkers

  • Functional studies may reveal oncogenic or tumor-suppressive roles

• Neurobiological disorders:

  • LRRC3C's participation in protein-protein interactions and signaling may impact neuronal function

  • Studies in neurodevelopmental and neurodegenerative contexts could identify novel disease mechanisms

  • Analysis of neural tissue expression patterns may identify region-specific functions

• Developmental biology:

  • Examining LRRC3C expression during embryonic development could reveal stage-specific roles

  • Knockout models may identify developmental processes requiring LRRC3C function

• Immunological processes:

  • LRRC3C's leucine-rich repeat domains share structural similarities with immune receptors

  • Investigation in immune cell populations may reveal roles in immune regulation

These research directions could provide insights into disease mechanisms and potentially identify LRRC3C as a therapeutic target.

What innovative techniques are being developed for LRRC3C protein analysis?

Emerging technologies offer new opportunities for LRRC3C research:

• Advanced imaging approaches:

  • Super-resolution microscopy for precise subcellular localization

  • Live-cell imaging with tagged LRRC3C to monitor dynamics

  • Correlative light and electron microscopy (CLEM) for structural context

• Proteomic techniques:

  • Proximity labeling methods (BioID, APEX) to map LRRC3C interaction networks

  • Cross-linking mass spectrometry to identify direct binding partners

  • Thermal proteome profiling to investigate LRRC3C stability and drug interactions

• Single-cell analysis:

  • Single-cell proteomics to examine LRRC3C expression heterogeneity

  • Spatial transcriptomics combined with protein validation to analyze expression patterns

• Structural biology approaches:

  • Cryo-EM analysis of LRRC3C complexes

  • Hydrogen-deuterium exchange mass spectrometry to probe conformational dynamics

These innovative methods can provide deeper insights into LRRC3C function beyond what traditional antibody-based approaches can achieve.

How might LRRC3C research contribute to therapeutic development?

LRRC3C research could impact therapeutic development through several avenues:

• Biomarker potential:

  • High-sensitivity ELISA assays (detecting as low as 0.066 ng/mL) could be developed for diagnostic or prognostic applications

  • Expression patterns in disease states might identify patient subgroups for targeted therapies

• Target validation:

  • Functional studies using antibodies and recombinant proteins can validate LRRC3C as a therapeutic target

  • Structure-function analysis could identify critical domains for drug targeting

• Therapeutic modalities:

  • If validated as a target, LRRC3C could be addressed through:

    • Blocking antibodies that disrupt specific interactions

    • Small molecule inhibitors targeting functional domains

    • Protein-protein interaction disruptors

    • RNA-based therapeutics to modulate expression

• Combination approaches:

  • Understanding LRRC3C's role in signaling networks could identify synergistic therapeutic combinations

  • Pathway analysis might reveal indirect approaches to modulate LRRC3C function

The continued development of research tools and methodologies for LRRC3C will be essential to realize these potential therapeutic applications.