GPRIN2 Antibody, Biotin conjugated

What is GPRIN2 and what is its biological function?

GPRIN2 (G protein-regulated inducer of neurite outgrowth 2) is a 458 amino acid protein primarily expressed in the cerebellum that plays a critical role in neurite outgrowth. It functions by interacting with activated G𝛼o and G𝛼i proteins to regulate neuronal development pathways. The gene encoding GPRIN2 is located on human chromosome 10q11.22. GPRIN2 has been identified in subcellular locations including the cytoplasm, nucleus, and as a secreted protein in the extracellular matrix, suggesting multiple functional roles . Alternative names for this protein include GRIN2, MGC15171, and mKIAA0514, which is important to recognize when reviewing literature across different research groups .

What applications is the biotin-conjugated GPRIN2 antibody suitable for?

The biotin-conjugated GPRIN2 polyclonal antibody is optimized for multiple experimental applications with specific recommended dilutions for each technique:

These applications allow researchers to detect and quantify GPRIN2 in various experimental contexts, from tissue localization studies to protein interaction assays . The biotin conjugation provides enhanced detection sensitivity when used with streptavidin or avidin detection systems compared to unconjugated antibodies.

How should the biotin-conjugated GPRIN2 antibody be stored to maintain optimal activity?

For maximum stability and reactivity, the biotin-conjugated GPRIN2 antibody should be stored at -20°C for up to 12 months . The antibody is supplied in an aqueous buffered solution containing 0.01M TBS (pH 7.4) with 1% BSA, 0.03% Proclin300, and 50% Glycerol, which helps maintain stability during freezing and thawing cycles . It is advisable to aliquot the antibody upon receipt to minimize repeated freeze-thaw cycles, which can degrade the protein and reduce its efficacy. When working with the antibody, thaw aliquots on ice and keep cold during experimental procedures to preserve the biotin conjugation and antibody binding capacity.

What species reactivity can be expected with the GPRIN2 antibody?

The biotin-conjugated GPRIN2 polyclonal antibody has demonstrated reactivity with multiple species:

The antibody was generated using a KLH-conjugated synthetic peptide derived from human GPRIN2 (immunogen range: 251-350/458), which explains its strong reactivity with human samples . Cross-reactivity with other species is based on sequence homology predictions, so validation experiments are recommended when using this antibody with non-human samples.

How does the biotin conjugation of GPRIN2 antibody enhance detection sensitivity in immunoassays?

The biotin conjugation of GPRIN2 antibody significantly enhances detection sensitivity through several biochemical mechanisms. Biotin forms an exceptionally strong non-covalent interaction with streptavidin/avidin (Kd ≈ 10^-15 M), which is one of the strongest non-covalent biological interactions known . This property allows for signal amplification through multivalent binding, as each streptavidin molecule can bind four biotin molecules.

In practical applications, this translates to:

• Enhanced signal detection: When using streptavidin-conjugated reporter enzymes (like HRP) or fluorophores, multiple reporter molecules can bind to each antibody molecule, amplifying the signal.

• Improved detection of low-abundance targets: The biotin-streptavidin system allows for detection of GPRIN2 even when present at very low concentrations in samples.

• Reduced background: The specificity of the biotin-streptavidin interaction minimizes non-specific binding compared to other detection systems.

This is particularly valuable in neuroscience research where GPRIN2 may be expressed at varying levels in different neuronal populations . For detection optimization, researchers should titrate both the biotin-conjugated primary antibody and the streptavidin-conjugated detection reagent to achieve the optimal signal-to-noise ratio.

What are the critical considerations for validating GPRIN2 antibody specificity in experimental designs?

Validating antibody specificity is crucial for generating reliable research data. For GPRIN2 biotin-conjugated antibodies, several validation strategies should be implemented:

• Peptide competition assays: Pre-incubate the antibody with its specific immunogen peptide (from the 251-350/458 region of human GPRIN2) before application to samples. This should abolish specific staining if the antibody is truly specific .

• Knockout/knockdown controls: Compare staining between GPRIN2 knockout/knockdown samples and wild-type samples to confirm specificity.

• Multiple antibody approach: Use different antibodies targeting distinct epitopes of GPRIN2 to confirm staining patterns.

• Cross-reactivity testing: When using the antibody across species, validate through Western blot analysis of tissue lysates from each species to confirm the antibody recognizes bands of the expected molecular weight (approximately 64 kDa for GPRIN2).

• Positive control tissue selection: Use cerebellum tissue for validation as GPRIN2 is known to be highly expressed in this brain region .

These validation steps are particularly important because GPRIN2 shares sequence homology with other GRIN family proteins, which could lead to cross-reactivity if not properly controlled for in experimental designs.

How can the biotin-conjugated GPRIN2 antibody be optimized for dual immunofluorescence studies?

Optimizing biotin-conjugated GPRIN2 antibody for dual immunofluorescence requires careful consideration of several methodological factors:

• Sequential detection protocol: Due to the strong biotin-streptavidin interaction, it's recommended to complete the biotin-streptavidin detection steps before introducing other antibodies. This typically involves:

  • Primary incubation with biotin-conjugated GPRIN2 antibody

  • Washing thoroughly

  • Incubation with fluorophore-conjugated streptavidin

  • Additional washing steps

  • Application of the second primary antibody (from a different host species)

  • Detection with species-specific secondary antibody

• Blocking endogenous biotin: Brain tissue contains endogenous biotin which can cause background staining. Use an avidin/biotin blocking kit before antibody application to minimize this issue .

• Compatible fluorophore selection: When selecting streptavidin-conjugated fluorophores, consider spectral overlap with other fluorophores used in the experiment. For example, if using FITC-based detection for a second target, avoid using fluorescein-related fluorophores with streptavidin to prevent cross-channel bleed-through .

• Concentration optimization: For dual staining, it may be necessary to reduce the concentration of the biotin-conjugated GPRIN2 antibody (consider starting at 1:400 for IHC-P) to minimize potential cross-reactivity or oversaturation of signal .

This approach allows for simultaneous visualization of GPRIN2 and other proteins of interest, particularly useful when studying GPRIN2's interaction with G proteins or neuronal markers in brain tissue sections.

What approaches can resolve contradictory results when using GPRIN2 antibody across different experimental platforms?

When faced with contradictory results using GPRIN2 antibody across different experimental platforms (e.g., IHC versus Western blot), systematic troubleshooting approaches should be employed:

• Epitope accessibility analysis: The biotin-conjugated GPRIN2 antibody targets an epitope in the 251-350 region of the 458 amino acid protein . This epitope may be differentially accessible in various applications due to protein folding, fixation effects, or denaturation conditions. Modifying sample preparation (e.g., different fixatives for IHC or denaturing conditions for Western blot) can help resolve these discrepancies.

• Cross-application validation strategy:

  • For contradictions between IHC and Western blot: Perform protein extraction from the same tissue used for IHC to directly compare expression levels.

  • For contradictions between in vitro and in vivo findings: Use primary neuronal cultures derived from the same species/strain used for in vivo studies.

• Isoform-specific detection: GPRIN2 may exist in multiple isoforms or undergo post-translational modifications that affect antibody recognition. Western blot analysis can identify if multiple bands are detected, suggesting the presence of isoforms, and mass spectrometry can be used for definitive identification .

• Sample preparation standardization: Different buffer compositions can affect epitope exposure:

  • For IHC: Compare heat-induced versus enzymatic antigen retrieval methods

  • For Western blot: Test both reducing and non-reducing conditions

  • For ELISA: Compare direct coating versus capture antibody approaches

This systematic approach can help reconcile apparently contradictory results and provide insight into the biological complexity of GPRIN2 expression and function across different experimental contexts.

How does the choice of detection system impact the sensitivity of biotin-conjugated GPRIN2 antibody assays?

The selection of an appropriate detection system significantly impacts assay sensitivity when using biotin-conjugated GPRIN2 antibodies. A comparative analysis of common detection systems reveals important differences:

NeutrAvidin-based detection systems often provide superior results for water-soluble targets due to reduced non-specific binding compared to standard avidin . This is particularly relevant for GPRIN2 studies in neural tissues where high lipid content can contribute to background noise. For quantitative applications like ELISA, enzymatic amplification using streptavidin-HRP provides better sensitivity than direct fluorescence methods, with detection limits potentially in the picogram range for GPRIN2 protein .

For microscopy applications, tyramide signal amplification (TSA) with biotin-conjugated GPRIN2 antibody can improve detection of low-abundance targets by generating multiple fluorophore depositions at the antibody binding site, significantly enhancing signal intensity while maintaining specificity.

What are the recommended controls for validating GPRIN2 localization in subcellular fractionation studies?

When conducting subcellular fractionation studies to investigate GPRIN2 localization, a comprehensive set of controls should be included to ensure accurate interpretation of results:

• Fraction purity markers: Include Western blot analysis of the following markers alongside GPRIN2 detection:

  Subcellular Compartment	Recommended Marker Proteins
  Cytoplasm	GAPDH, β-tubulin
  Nucleus	Lamin B1, Histone H3
  Membrane	Na+/K+ ATPase, Cadherin
  Secreted/ECM	Fibronectin, Collagen

  This is particularly important as GPRIN2 has been reported in multiple subcellular locations including cytoplasm, nucleus, and as a secreted protein .

• Crossover contamination assessment: Quantify the presence of compartment-specific markers in each fraction to mathematically correct for contamination between fractions.

• Co-fractionation studies: Include known GPRIN2 interacting partners such as G𝛼o and G𝛼i proteins to confirm functional associations in specific compartments .

• Differential extraction controls: Compare results from differential detergent extraction methods (e.g., Triton X-100 vs. digitonin) which can differentially extract GPRIN2 from distinct membrane microdomains.

• Physiological state comparisons: Analyze GPRIN2 distribution under different cellular conditions (e.g., resting vs. stimulated neurons) to assess dynamic relocalization.

These controls help distinguish true subcellular localization from fractionation artifacts and provide insights into the functional significance of GPRIN2 in different cellular compartments.

How should researchers optimize antigen retrieval for GPRIN2 detection in fixed neural tissues?

Antigen retrieval optimization is critical for successful detection of GPRIN2 in fixed neural tissues, particularly when using biotin-conjugated antibodies. The following methodological approach addresses the unique challenges of neural tissue immunohistochemistry:

• Comparative evaluation of retrieval methods:

  Retrieval Method	Buffer Composition	Conditions	Best For
  Heat-Induced (HIER)	10mM Citrate buffer (pH 6.0)	95°C, 20 minutes	Formalin-fixed tissues with moderate fixation
  Heat-Induced (HIER)	10mM Tris-EDTA (pH 9.0)	95°C, 30 minutes	Heavily fixed tissues, enhanced GPRIN2 detection
  Enzymatic	Proteinase K (10μg/ml)	37°C, 10 minutes	Fresh-frozen sections, light fixation
  Combined	Proteinase K followed by HIER	Sequential application	Difficult samples with high lipid content

  Based on protocols for similar neural proteins and antibody applications, Tris-EDTA (pH 9.0) buffer often provides superior retrieval for antibodies like biotin-conjugated GPRIN2 in neural tissues .

• Fixation-specific considerations: For perfusion-fixed brain tissues, extend HIER treatment time by 5-10 minutes compared to immersion-fixed samples.

• Regional optimization: Cerebellar tissue, where GPRIN2 is highly expressed, may require gentler retrieval conditions compared to cortical regions to preserve morphology while achieving optimal antigen exposure .

• Microwave vs. pressure cooker comparison: Pressure cooker methods often yield more consistent results for neural tissue antigens, but should be empirically compared for GPRIN2 detection specifically.

• Post-retrieval cooling protocol: Allow slides to cool slowly to room temperature (approximately 20 minutes) after HIER to minimize tissue detachment and preserve morphology before proceeding with antibody incubation.

These optimized retrieval approaches significantly enhance GPRIN2 detection sensitivity while maintaining tissue integrity, critical for accurate localization studies in neural tissues.

How can researchers differentiate between specific GPRIN2 signals and non-specific biotin-related background in neural tissues?

Distinguishing specific GPRIN2 signals from biotin-related background is a critical challenge in neural tissue analysis. Several methodological approaches can address this issue:

• Endogenous biotin blocking strategy: Brain tissue contains significant endogenous biotin, particularly in regions like the hippocampus and cerebellum where GPRIN2 is also expressed. Implement a sequential blocking protocol:

  • Pre-block with unconjugated avidin (10-15 minutes)

  • Wash briefly

  • Block with excess biotin (10-15 minutes)

  • Proceed with normal blocking buffer
    This approach saturates endogenous biotin and biotin-binding sites before introducing the biotin-conjugated GPRIN2 antibody .

• Comparative analysis with alternative detection methods: Validate critical findings using a non-biotin detection system, such as directly conjugated fluorescent antibodies or HRP-conjugated secondary antibodies.

• Tissue-specific negative controls: Include sections from tissues known to be negative for GPRIN2 but containing endogenous biotin (e.g., liver sections) to establish baseline biotin-related background levels.

• Streptavidin control: Include control sections treated with streptavidin-reporter systems but no primary antibody to visualize endogenous biotin patterns.

• Pattern analysis: GPRIN2 shows characteristic subcellular distribution patterns (cytoplasmic, nuclear, and extracellular matrix) that can be distinguished from the punctate mitochondrial pattern typical of endogenous biotin .

Implementing these controls and analytical approaches allows for confident identification of true GPRIN2 signals versus non-specific background, particularly important in quantitative studies of GPRIN2 expression in neural tissues.

What approaches can resolve data discrepancies between GPRIN2 detection methods in complex neural samples?

When faced with discrepancies in GPRIN2 detection across different methods, a systematic troubleshooting approach can help resolve inconsistencies:

• Epitope mapping assessment: The biotin-conjugated GPRIN2 antibody targets a specific epitope (region 251-350/458) that may be differentially accessible depending on the method . Consider:

  • Native vs. denatured conditions affecting epitope exposure

  • Fixation-induced epitope masking in IHC vs. Western blot

  • Protein-protein interactions potentially blocking epitope access in co-IP experiments

• Cross-validating quantification methods:

  Method	Strengths	Limitations	Best Practices
  Western Blot	Protein size verification	Semi-quantitative	Include recombinant GPRIN2 standards
  IHC/IF	Spatial localization	Subjective quantification	Use automated image analysis
  ELISA	Precise quantification	No size verification	Multiple antibody approach
  Mass Spectrometry	Definitive identification	Complex sample prep	Target specific peptides in 251-350 region

• Integrated validation workflow: For critical findings, implement a sequential validation approach:

  • Initial discovery with biotin-conjugated antibody

  • Confirmation with unconjugated GPRIN2 antibody

  • Orthogonal validation with mRNA analysis (ISH or RT-PCR)

  • Final verification with genetic approaches (knockdown/knockout)

• Statistical approach to discrepancies: When methods yield different results, consider:

  • Bland-Altman analysis to quantify systematic differences between methods

  • Multiple regression models incorporating method-specific variables

  • Meta-analysis approaches when combining data across multiple detection systems

This comprehensive approach helps resolve apparent contradictions by identifying method-specific biases and establishing consensus findings with higher confidence.

What statistical approaches are most appropriate for analyzing GPRIN2 expression data from biotin-amplified detection systems?

Biotin-amplified detection systems for GPRIN2 present unique statistical challenges due to their non-linear signal amplification properties. The following statistical approaches are recommended:

• Transformation for normality: Biotin-streptavidin amplified signals often follow log-normal rather than normal distributions. Apply logarithmic transformation before parametric statistical analyses to meet assumptions of normality.

• Calibration curve modeling:

  Signal Range	Recommended Model	Application
  Low to medium	Linear regression	Standard ELISA
  Full range	Four-parameter logistic (4PL)	High sensitivity assays
  With hook effect	Five-parameter logistic (5PL)	Complex neural lysates

  The 4PL or 5PL models account for the sigmoidal dose-response relationship often observed with biotin-amplified detection systems, providing more accurate quantification across a wider dynamic range .

• Appropriate reference standard approach: For relative quantification, implement:

  • Recombinant GPRIN2 protein standards spanning physiological concentration ranges

  • Housekeeping protein normalization with careful selection of reference proteins stable in neural tissues

  • MIQE-compliant protocols when using qPCR to correlate with protein expression

• Statistical methods for grouped data:

  • For normally distributed transformed data: ANOVA with post-hoc tests

  • For non-parametric approaches: Kruskal-Wallis with Dunn's post-hoc test

  • For longitudinal studies: Mixed-effects models accounting for repeated measures

• Signal saturation correction: Implement centering-and-scaling transformations or hyperbolic regression models to correct for signal saturation in highly expressed samples.

These statistical approaches ensure accurate quantification and comparison of GPRIN2 expression across experimental conditions, particularly important when using highly sensitive biotin-amplified detection systems that can otherwise lead to misleading results if conventional statistical methods are applied without consideration of their specific signal characteristics.

How can biotin-conjugated GPRIN2 antibodies be used to investigate neurite outgrowth mechanisms?

Biotin-conjugated GPRIN2 antibodies offer powerful tools for investigating neurite outgrowth mechanisms through several advanced experimental approaches:

• Time-course visualization protocols: The biotin-streptavidin system provides exceptional sensitivity for tracking GPRIN2 dynamics during neurite outgrowth:

  • Fix neuronal cultures at defined time points (6h, 12h, 24h, 48h, 72h)

  • Process with biotin-conjugated GPRIN2 antibody (1:200 dilution)

  • Visualize with fluorescent streptavidin

  • Co-stain with cytoskeletal markers (β-III-tubulin) and growth cone markers (GAP-43)

  This approach reveals the spatial-temporal relationship between GPRIN2 expression/localization and neurite extension phases .

• G-protein interaction studies: GPRIN2 functions through interactions with G𝛼o and G𝛼i proteins. Implement:

  • Proximity ligation assays (PLA) using biotin-conjugated GPRIN2 antibody and antibodies against G𝛼o/G𝛼i

  • Co-immunoprecipitation followed by Western blot analysis

  • FRET-based interaction studies in live neurons

  These approaches reveal when and where GPRIN2-G protein interactions occur during neurite outgrowth .

• Quantitative morphometric analysis workflow:

  • Culture neurons on gridded coverslips

  • Treat with modulators of G-protein signaling

  • Immunostain with biotin-GPRIN2 antibody and neurite markers

  • Acquire high-resolution images

  • Perform automated Sholl analysis and neurite tracing

  • Correlate GPRIN2 expression levels with neurite complexity metrics

• Molecular perturbation studies: Combine GPRIN2 overexpression or knockdown with:

  • Live-cell imaging of neurite dynamics

  • Quantification of growth cone turning responses

  • Assessment of branch formation frequency

  • Analysis of neurite stabilization/retraction events

These methodologies leverage the sensitivity and specificity of biotin-conjugated GPRIN2 antibodies to reveal the functional role of this protein in neurodevelopmental processes, providing insights into both normal development and potential pathological mechanisms in neurodevelopmental disorders.

What considerations are important when using GPRIN2 antibodies to study potential links to neurological disorders?

When investigating GPRIN2's potential involvement in neurological disorders, several critical methodological considerations must be addressed:

• Case-control tissue selection strategy:

  • Match cases and controls for age, sex, post-mortem interval, and brain region

  • Consider comorbidities that could affect GPRIN2 expression

  • Establish standardized tissue processing protocols to minimize technical variability

  • Include samples from multiple brain regions since GPRIN2 is differentially expressed across neural tissues

• Disorder-specific protocol modifications:

  Neurological Condition	Special Considerations	Recommended Approaches
  Neurodevelopmental disorders	Developmental timepoint analysis	Compare GPRIN2 expression across developmental stages
  Neurodegenerative diseases	Protein aggregation interference	Use specialized extraction buffers; compare soluble vs. insoluble fractions
  Neuropsychiatric conditions	Regional specificity	Layer-specific analysis in cortical regions
  Traumatic brain injury	Acute vs. chronic changes	Time-course studies with consistent sampling locations

• Integration with genetic data: Correlate GPRIN2 protein expression patterns with:

  • Known genetic variants in GPRIN2 or interacting partners

  • Transcriptomic profiles from the same samples

  • Pathway analysis incorporating G-protein signaling components

• Functional validation requirements:

  • Develop in vitro models recapitulating disorder-relevant phenotypes

  • Establish causality through genetic manipulation (CRISPR/Cas9)

  • Validate findings across multiple model systems (cell lines, primary cultures, animal models)

  • Test pharmacological modulators of GPRIN2-G protein interactions

• Technical controls for pathological samples:

  • Test for fixation artifacts in post-mortem tissues

  • Validate antibody specificity in disease-state tissues

  • Account for disease-associated changes in reference genes/proteins

These methodological considerations ensure that findings regarding GPRIN2's role in neurological disorders are robust, reproducible, and biologically meaningful, potentially leading to new insights into disease mechanisms or therapeutic targets.

What strategies can resolve weak or absent signals when using biotin-conjugated GPRIN2 antibody in Western blotting?

When encountering weak or absent signals with biotin-conjugated GPRIN2 antibody in Western blotting, implement this systematic troubleshooting approach:

• Sample preparation optimization:

  • Use fresh RIPA buffer supplemented with protease inhibitors

  • For neural tissues, add phosphatase inhibitors to preserve potential phosphorylated forms of GPRIN2

  • Try gentler lysis methods as harsh detergents may disrupt epitopes in the 251-350 region

  • Include membrane-enrichment steps as GPRIN2 can associate with membrane fractions

• Protein transfer enhancement protocol:

  Transfer Parameter	Standard Protocol	Optimized Protocol for GPRIN2
  Transfer time	1 hour	16 hours (overnight)
  Buffer composition	Standard Towbin	Towbin + 0.05% SDS
  Current	Constant mA	Sequential current reduction
  Membrane type	PVDF	PVDF with 0.2μm pore size

  These modifications enhance transfer efficiency of GPRIN2, particularly important for membrane-associated proteins .

• Signal amplification strategies:

  • Use enhanced chemiluminescence (ECL) substrates specifically designed for biotin-streptavidin systems

  • Implement tyramide signal amplification (TSA)

  • Consider poly-HRP streptavidin conjugates for multiplicative signal enhancement

  • Optimize incubation temperature (try 4°C overnight) to increase specific binding

• Blocking optimization:

  • Test alternative blocking agents (BSA vs. non-fat milk vs. commercial blockers)

  • Reduce blocking stringency if GPRIN2 expression is low

  • Implement specialized blocking for biotin-streptavidin systems

  • Consider adding 0.05% Tween-20 to reduce background while maintaining specific signals

• Antibody and detection reagent titration matrix:

  • Test wider concentration ranges (1:100 to 1:5000) than standard protocols suggest

  • Create a dilution matrix crossing primary antibody with streptavidin-HRP dilutions

  • Extend primary antibody incubation times (up to 48 hours at 4°C for low abundance targets)

These optimized protocols significantly enhance detection sensitivity for GPRIN2 in Western blotting applications, particularly important when working with samples where GPRIN2 expression may be naturally low or modified in experimental conditions.

How can researchers minimize background and optimize signal-to-noise ratio in IHC with biotin-conjugated GPRIN2 antibody?

Achieving optimal signal-to-noise ratio in IHC with biotin-conjugated GPRIN2 antibody requires a comprehensive approach to background reduction:

• Endogenous biotin blocking protocol optimization:

  • Implement sequential avidin-biotin blocking (15 minutes each)

  • For neural tissues with high biotin content, consider extending blocking to 30 minutes

  • Pre-incubate sections with unconjugated streptavidin prior to primary antibody

  • Add 1mM biotin to antibody diluent buffer to continuously compete with endogenous biotin

• Tissue-specific background reduction strategies:

  Tissue Type	Background Source	Reduction Strategy
  Brain tissue	Lipofuscin autofluorescence	Add Sudan Black B (0.1%) post-staining
  Fixed neural tissue	Aldehyde-induced autofluorescence	Pretreat with sodium borohydride (0.1%)
  Aged brain samples	Cross-linked lipoproteins	Add 0.3% Triton X-100 to antibody diluent
  Peripheral nerve	Myelin components	Use saponin instead of Triton X-100

• Antibody diluent optimization:

  • Add 5% normal serum from the species of the secondary reagent

  • Include 0.1% carrier proteins like BSA or casein

  • Add 0.05-0.3% Triton X-100 for permeabilization

  • Consider adding 5% non-fat milk to further reduce non-specific binding

• Signal-to-noise enhancement workflow:

  • Titrate primary antibody with narrower dilution ranges around optimal point (start with 1:200-1:400)

  • Reduce streptavidin-conjugate concentration to minimize background

  • Implement thorough washing (5 washes of 5 minutes each between reagents)

  • Consider using TSA amplification only for very low abundance targets

• Advanced counterstaining protocol:

  • Use fluorescent nuclear counterstains at reduced concentrations

  • Implement sequential imaging with careful exposure settings

  • Consider spectral unmixing for overlapping fluorophores

  • Use reference spectra libraries for tissue autofluorescence subtraction

These optimized approaches significantly improve signal-to-noise ratio in IHC applications of biotin-conjugated GPRIN2 antibody, enabling reliable detection and accurate localization even in challenging neural tissue samples.