SRR Antibody, FITC conjugated

What is SRR Antibody, FITC conjugated and what biological systems can it be used in?

SRR Antibody, FITC conjugated is a polyclonal antibody developed in rabbits that specifically targets human serine racemase (SRR), an enzyme that catalyzes the synthesis of D-serine from L-serine. The antibody comes with a fluorescein isothiocyanate (FITC) conjugation, which enables direct fluorescent detection in various applications. According to the product specifications, this particular antibody has been tested and validated for human samples . SRR plays a crucial role in neuroscience research as it produces D-serine, which functions as a key coagonist with glutamate at NMDA receptors, making this antibody particularly valuable for neurological studies . The antibody's IgG isotype and FITC conjugation make it suitable for direct detection without the need for secondary antibodies in applications like flow cytometry, immunofluorescence microscopy, and other fluorescence-based detection methods. Researchers should note that while this antibody is primarily validated for human samples, cross-reactivity with other species should be empirically determined before use in non-human experimental systems.

How does serine racemase function in neural pathways and why is it important to study?

Serine racemase (SRR) performs the critical enzymatic conversion of L-serine to D-serine, which serves as an essential coagonist with glutamate at NMDA receptors in neural tissues . Beyond this primary function, SRR also exhibits dehydratase activity toward both L-serine and D-serine, adding complexity to its metabolic role in neural systems . The enzyme's activity directly impacts glutamatergic neurotransmission by regulating available D-serine levels, which can affect synaptic plasticity, learning, memory formation, and other higher cognitive functions. Dysregulation of SRR has been implicated in several neurological and psychiatric conditions, including schizophrenia, epilepsy, neurodegenerative diseases, and certain forms of cognitive impairment. Given its alternative nomenclature as "D-serine ammonia-lyase" or "D-serine dehydratase," researchers should be aware of these synonyms when reviewing literature . Studying SRR using tools like FITC-conjugated antibodies allows visualization of its expression patterns, subcellular localization, and potential changes in pathological states, contributing to our understanding of excitatory neurotransmission mechanisms and potential therapeutic targets.

What are the basic storage and handling recommendations for maintaining SRR Antibody, FITC conjugated activity?

Proper storage and handling of SRR Antibody, FITC conjugated is essential for maintaining its specificity and fluorescent signal integrity. Based on standard antibody protocols similar to other FITC-conjugated antibodies, these reagents should be stored at -20°C or -80°C for long-term preservation as indicated in the product specifications . When working with the antibody, it's important to minimize exposure to light due to the light-sensitive nature of the FITC fluorophore, which can photobleach with prolonged light exposure. Repeated freeze-thaw cycles should be avoided as they can lead to protein denaturation and degradation of antibody performance, similar to recommendations for other antibody products . For reconstituted antibodies, short-term storage at 2-8°C is typically acceptable for up to one month under sterile conditions, while for longer periods (up to 6 months), aliquoting and returning to -20°C or -70°C is recommended . When preparing working dilutions, researchers should use buffers that maintain protein stability, typically PBS with a carrier protein such as BSA (0.1-1%) and possibly a preservative like sodium azide (though caution is needed if using in live cell applications). As with all immunological reagents, validation of each new lot is advisable to ensure consistent performance in your specific experimental system.

How should I design flow cytometry experiments with SRR Antibody, FITC conjugated for optimal results?

When designing flow cytometry experiments with SRR Antibody, FITC conjugated, researchers should first determine the optimal antibody concentration through titration experiments. While specific recommendations for the SRR antibody aren't provided in the sources, a general starting point for FITC-conjugated antibodies in flow cytometry is approximately 0.25 μg per test, as suggested for similar conjugated antibodies . The FITC fluorophore has excitation/emission peaks at approximately 495/519 nm, requiring appropriate laser and filter settings (typically 488 nm excitation laser and 530/30 nm bandpass filter). For intracellular targets like SRR, a permeabilization step is crucial—typically using reagents like saponin, Triton X-100, or commercial permeabilization buffers—since SRR functions inside cells synthesizing D-serine from L-serine . Proper compensation is essential when designing multicolor panels, as FITC has spectral overlap with other fluorophores like PE. Include appropriate controls in your experimental design: unstained cells, isotype controls (Rabbit IgG-FITC), fluorescence-minus-one (FMO) controls, and positive controls (cells known to express SRR). For analyzing neurons or glial cells expressing SRR, consider using gentle dissociation methods to maintain cellular integrity and epitope accessibility. When analyzing data, use appropriate gating strategies to exclude dead cells, debris, and doublets before assessing SRR expression levels.

What immunofluorescence protocols work best for visualizing SRR localization in neural tissue sections?

For optimal visualization of SRR localization in neural tissue sections using FITC-conjugated SRR antibody, begin with properly fixed tissue—4% paraformaldehyde is typically suitable for preserving both tissue architecture and antigenicity. When working with frozen sections, a dilution range of 1:400 to 1:800 is generally appropriate for FITC-conjugated antibodies, though optimization may be necessary for SRR antibody specifically . The protocol should include sufficient permeabilization steps (0.1-0.3% Triton X-100 in PBS for 10-20 minutes) to allow antibody access to intracellular SRR, followed by a blocking step using 5-10% normal serum from a species different from the antibody host (non-rabbit serum in this case) with 1% BSA in PBS to minimize non-specific binding. For co-localization studies, consider pairing the SRR-FITC antibody with markers for specific neural cell types (neurons, astrocytes, or microglia) or subcellular compartments using compatible fluorophores that don't overlap with FITC's emission spectrum. Counterstaining with DAPI (excitation/emission: 358/461 nm) allows nuclear visualization without interfering with the FITC signal. To control for autofluorescence, which is common in neural tissues, include sections stained with isotype control antibodies and consider using Sudan Black B treatment (0.1-1% in 70% ethanol) to quench lipofuscin autofluorescence. For quantitative analysis of SRR expression patterns across different brain regions or in disease models, use consistent exposure settings and analysis parameters to enable valid comparisons between experimental groups.

What approaches can be used to validate SRR antibody specificity for ensuring reliable experimental results?

Validating the specificity of SRR Antibody, FITC conjugated is crucial for generating reliable experimental data and avoiding misinterpretation of results. The gold standard approach involves comparing staining patterns between wild-type samples and SRR knockout models, where the absence of signal in knockout tissues would confirm specificity. Another powerful validation method is performing peptide competition assays, where pre-incubating the antibody with purified recombinant SRR protein (such as the Recombinant Human Serine racemase mentioned in the search results) should abolish or significantly reduce the staining signal if the antibody is truly specific . For human samples where genetic models aren't available, RNA interference techniques (siRNA or shRNA) to knockdown SRR expression provides an alternative validation approach, with reduced staining intensity expected in knockdown samples. Western blot analysis should show a single band at the expected molecular weight for SRR (approximately 37 kDa), and correlation between protein and mRNA expression levels across different tissues or experimental conditions adds another layer of validation. Side-by-side comparison with other validated antibodies targeting SRR from different host species or recognizing different epitopes should show consistent staining patterns. Additionally, for FITC-conjugated antibodies specifically, researchers should include controls to distinguish between specific binding and potential non-specific fluorescence, such as isotype controls (Rabbit IgG-FITC) and testing the unconjugated primary antibody followed by a secondary detection system.

How can SRR Antibody, FITC conjugated be utilized to study the relationship between D-serine regulation and NMDA receptor function?

The SRR Antibody, FITC conjugated offers a powerful tool for investigating the spatial and temporal relationship between serine racemase expression and NMDA receptor function in neural tissues. Researchers can employ this antibody in dual immunofluorescence studies alongside NMDA receptor subunit antibodies (with compatible fluorophores) to visualize their spatial proximity and potential co-regulation in both physiological and pathological states. In brain slice preparations, combining immunofluorescence with electrophysiological recordings allows correlation between SRR expression levels (detected via the FITC signal) and NMDA receptor-dependent currents in the same neuronal populations. For in vitro studies, researchers can use the antibody to track changes in SRR expression following pharmacological manipulation of NMDA receptor activity (using agonists like NMDA or antagonists like AP5/MK-801), providing insights into feedback regulation between these systems. The antibody can also be applied in flow cytometry to quantify SRR expression in isolated neural cells from various brain regions, enabling correlation with region-specific D-serine levels and NMDA receptor functionality. Additionally, the FITC-conjugated SRR antibody can be employed in time-course studies following neural activity induction (using models like long-term potentiation) to track dynamic changes in SRR expression or subcellular localization that may support activity-dependent D-serine production for NMDA receptor modulation. These approaches collectively help elucidate the mechanistic links between SRR activity, D-serine availability, and glutamatergic neurotransmission in both normal physiology and neurological disorders.

What techniques can combine SRR antibody detection with functional assessment of D-serine production?

Integrating SRR antibody detection with functional assessment of D-serine production creates a powerful approach for understanding the relationship between enzyme expression and activity. Researchers can employ microdialysis in defined brain regions followed by high-performance liquid chromatography (HPLC) to quantify extracellular D-serine levels, then perform post-hoc immunofluorescence with the FITC-conjugated SRR antibody on the same tissue to correlate D-serine release with SRR expression patterns. Another sophisticated approach involves using biosensor technology, where microelectrodes coated with D-amino acid oxidase can detect real-time D-serine release in brain slices, which can then be fixed and stained with SRR-FITC antibody to correlate functional output with enzyme localization. For cell culture systems, media sampling for D-serine quantification (using HPLC or enzymatic assays) can be paired with flow cytometric analysis of SRR expression using the FITC-conjugated antibody in the same cell populations. Since SRR has both racemase and dehydratase activities toward L-serine and D-serine as noted in the product description, researchers might employ isotope-labeled serine substrates to track metabolic conversion while simultaneously monitoring SRR expression levels with the antibody . Additionally, calcium imaging to assess NMDA receptor activation (which requires D-serine as a co-agonist) can be combined with post-hoc SRR immunofluorescence to establish functional relationships between SRR expression, D-serine availability, and downstream receptor activation in neural circuits. These multimodal approaches provide mechanistic insights beyond what either technique alone could offer.

How can SRR Antibody, FITC conjugated be used to investigate neuroinflammatory conditions and their impact on D-serine signaling?

The SRR Antibody, FITC conjugated provides a valuable tool for investigating how neuroinflammatory processes affect D-serine signaling pathways in various neuropathological conditions. Researchers can employ this antibody in immunofluorescence studies of brain tissue from neuroinflammatory disease models or patient samples, co-staining with markers for microglia (Iba1), astrocytes (GFAP), and inflammatory cytokines to assess how inflammation alters SRR expression patterns in specific cell types. Flow cytometric analysis using the FITC-conjugated SRR antibody can quantify changes in SRR protein levels within isolated glial cell populations after experimental inflammatory stimulation (such as LPS treatment or cytokine exposure), providing insights into how inflammatory mediators regulate this enzyme. Time-course studies following inflammatory insults can reveal the temporal dynamics of SRR expression changes relative to inflammatory marker upregulation and resolution, helping to establish cause-effect relationships in neuroinflammatory cascades. The antibody can additionally be utilized in models of specific neurological disorders with inflammatory components (such as multiple sclerosis, Alzheimer's disease, or ischemic stroke) to determine how disease-specific inflammatory processes impact the D-serine synthesis pathway. For translational relevance, researchers might correlate SRR expression changes detected via this antibody with functional outcomes in behavioral tests of cognitive or motor function in disease models, potentially identifying therapeutic windows where modulating D-serine levels might counteract inflammation-induced neural dysfunction. These approaches collectively contribute to our understanding of how neuroinflammation affects glutamatergic neurotransmission through alterations in D-serine signaling pathways.

What are the most common problems encountered when using FITC-conjugated antibodies in neural tissue, and how can they be overcome?

When working with FITC-conjugated antibodies like SRR Antibody in neural tissue, researchers frequently encounter autofluorescence challenges that can mask specific signals. Neural tissues contain lipofuscin pigments and other autofluorescent compounds that emit in the same spectral range as FITC. This issue can be mitigated through several approaches: treatment with Sudan Black B (0.1-0.3% in 70% ethanol for 5-10 minutes after immunostaining), application of copper sulfate solution (1-5 mM CuSO₄ in 50 mM ammonium acetate), or using commercially available autofluorescence quenchers specifically designed for neural tissues. Another common problem is photobleaching of the FITC fluorophore during imaging, resulting in signal loss over time. To address this, researchers should minimize exposure to excitation light, add anti-fade agents to mounting media, and consider acquiring images from less-exposed areas of the sample first . The sensitivity limitations of FITC, particularly for low-abundance targets like SRR in certain neural populations, may necessitate signal amplification methods such as tyramide signal amplification or using anti-FITC antibodies conjugated to brighter fluorophores, as mentioned in the background information on FITC . For aged or long-fixed tissue samples, antigen retrieval methods may be essential for exposing the SRR epitope, typically using citrate buffer (pH 6.0) or EDTA buffer (pH 8.0) with controlled heating. Additionally, penetration issues in thick neural tissue sections can be addressed by extending incubation times, increasing detergent concentration in buffers, or employing specialized clearing techniques like CLARITY or iDISCO that maintain fluorophore integrity while enhancing antibody penetration.

How can researchers optimize signal-to-noise ratio when using SRR Antibody, FITC conjugated in complex neural samples?

Optimizing signal-to-noise ratio with SRR Antibody, FITC conjugated in complex neural samples requires a multifaceted approach addressing several technical aspects. First, titration of antibody concentration is essential—while specific optimal dilutions for this particular antibody aren't provided in the search results, starting with manufacturer recommendations and testing a dilution series (e.g., 1:200, 1:400, 1:800) helps identify the concentration that maximizes specific signal while minimizing background . The blocking step is crucial for neural tissues; use 5-10% normal serum (from a species different than the host species of the antibody) with 1% BSA and 0.1-0.3% Triton X-100 in PBS for 1-2 hours at room temperature to reduce non-specific binding. Sample preparation significantly impacts background; perfusion-fixed tissues generally yield cleaner results than immersion-fixed samples, and fresh frozen sections often provide better signal-to-noise ratios than paraffin-embedded sections for fluorescence applications. For confocal microscopy of FITC-labeled samples, optimize pinhole settings to exclude out-of-focus light, and adjust laser power and detector gain to capture specific signal while avoiding autofluorescence. Consider spectral unmixing techniques if your imaging system supports them, allowing separation of FITC signal from autofluorescence based on their distinct spectral properties. For quantitative studies, always image experimental and control samples under identical acquisition settings, and consider using software-based background subtraction methods with carefully selected reference regions. Additionally, applying a post-staining wash with high-salt PBS (containing 0.5-1.0 M NaCl) can help reduce non-specific electrostatic interactions between the antibody and tissue components, thereby enhancing signal specificity.

What validation controls should be included when examining potential cross-reactivity of SRR Antibody with related enzymes?

When examining potential cross-reactivity of SRR Antibody, FITC conjugated with related enzymes, researchers should implement a comprehensive set of validation controls to ensure signal specificity. First, include appropriate negative controls by testing the antibody on tissues or cells known to lack SRR expression, as well as on SRR knockout models if available, where any signal would indicate cross-reactivity. Peptide competition assays provide another crucial control—pre-incubating the antibody with purified recombinant SRR protein should eliminate specific staining, while pre-incubation with structurally related enzymes (such as serine dehydratase or other PLP-dependent enzymes) should not affect the signal if the antibody is truly specific . Western blot analysis comparing purified preparations of SRR and related enzymes can provide biochemical evidence of specificity, with the antibody showing reactivity only to SRR at its expected molecular weight. For cell-based assays, create expression systems with controlled overexpression of SRR and structurally similar enzymes tagged with distinct fluorescent proteins (e.g., RFP), then stain with the FITC-conjugated SRR antibody—overlap should occur only with the SRR-expressing population. Comparative staining with alternative antibodies targeting SRR through different epitopes offers additional validation, as consistent staining patterns across different antibodies increase confidence in specificity. Researchers should also perform sequence alignment analyses to identify regions of homology between SRR and related enzymes, which helps predict potential cross-reactivity based on epitope conservation. Finally, include concentration gradients in validation experiments, as cross-reactivity often manifests at higher antibody concentrations, allowing determination of a working concentration range that maximizes specificity.

How should researchers interpret variable SRR expression patterns across different neural cell types?

When interpreting variable SRR expression patterns across different neural cell types using FITC-conjugated SRR antibody, researchers should consider both biological significance and technical factors. Biologically, differential expression of SRR between neurons, astrocytes, and microglia likely reflects cell type-specific roles in D-serine metabolism and NMDA receptor modulation. Neurons traditionally utilize D-serine as a co-agonist for NMDA receptors, while astrocytes have been implicated in both the production and release of D-serine, suggesting potential for region-specific and activity-dependent expression patterns . Quantitative analysis should include measurement of both signal intensity (indicating expression level per cell) and the proportion of positive cells within each population, ideally using standardized approaches like mean fluorescence intensity (MFI) for flow cytometry or integrated density measurements for microscopy. When comparing expression across brain regions or in disease states, normalize data to appropriate reference regions or control samples while accounting for regional variations in tissue autofluorescence. For developmental studies, establish baseline expression trajectories in control animals before interpreting disease-related changes. From a technical perspective, validation through multiple techniques strengthens interpretation—findings from immunofluorescence should be corroborated with complementary methods like in situ hybridization for SRR mRNA, Western blotting of isolated cell populations, or single-cell RNA sequencing data. When observing unexpected expression patterns, consider potential post-translational modifications or protein-protein interactions that might mask antibody epitopes in specific cellular contexts. Additionally, correlate expression patterns with functional data on D-serine levels and NMDA receptor activity to establish the physiological relevance of observed expression differences across neural cell types.

What insights can comparative studies of SRR expression in healthy versus pathological neural tissues provide?

Comparative studies of SRR expression in healthy versus pathological neural tissues using FITC-conjugated SRR antibody can yield valuable insights into disease mechanisms and potential therapeutic approaches. In neurodegenerative conditions like Alzheimer's disease, alterations in SRR expression may correlate with changes in synaptic plasticity and cognitive decline, potentially linking D-serine metabolism dysfunction to disease progression. When designing such comparative studies, researchers should employ quantitative approaches—utilizing consistent imaging parameters, automated analysis algorithms, and appropriate statistical methods—to detect subtle but significant changes in SRR expression levels or subcellular distribution patterns. Co-staining with markers of disease pathology (such as amyloid plaques, neurofibrillary tangles, or inflammatory markers) can reveal spatial relationships between SRR expression changes and pathological features. Temporal analysis across disease progression stages provides insights into whether SRR alterations represent early disease biomarkers or consequential changes following pathology onset. For human tissue studies, careful matching of control and pathological samples for age, post-mortem interval, and brain region is essential for valid comparisons, as these factors can independently affect protein expression and preservation. Researchers should correlate SRR expression changes with functional outcomes where possible, such as electrophysiological measures of NMDA receptor activity or behavioral assessments in animal models. When interpreting decreased SRR immunoreactivity in disease states, distinguish between reduced expression, protein degradation, or epitope masking through additional validation approaches such as mRNA analysis or alternative antibodies. The translational potential of such comparative studies lies in identifying whether restoring normal SRR expression patterns or enhancing D-serine production could represent viable therapeutic strategies for conditions involving glutamatergic signaling dysfunction.

How can SRR antibody studies be integrated with functional NMDA receptor assays to advance neuroscience research?

Integrating SRR antibody studies with functional NMDA receptor assays creates a powerful approach for understanding D-serine signaling in complex neural circuits. Researchers can combine patch-clamp electrophysiology with post-hoc immunofluorescence using FITC-conjugated SRR antibody to correlate NMDA receptor-mediated currents with SRR expression levels in the same neurons or in adjacent glial cells, providing insights into local D-serine production and utilization. Calcium imaging using NMDA receptor-specific calcium indicators (like GCaMP with pharmacological isolation) can be performed on live tissue, followed by fixation and SRR immunostaining to map the relationship between receptor activation patterns and enzyme distribution. In brain slice preparations, researchers can manipulate SRR activity through genetic or pharmacological approaches, then assess the resulting changes in both NMDA receptor function (using electrophysiology or calcium imaging) and SRR expression/localization (using the FITC-conjugated antibody). For behavioral studies, tissue collected after cognitive or memory tasks can be analyzed for SRR expression patterns in relation to task performance and NMDA receptor-dependent plasticity markers. The technological integration can extend to in vivo approaches, where fiber photometry or miniscope imaging of NMDA receptor activity (using genetically encoded indicators) can be combined with post-experiment tissue analysis of SRR distribution using the FITC-conjugated antibody. When analyzing such integrated datasets, multivariate statistical approaches and computational modeling help elucidate the complex relationships between enzyme expression, D-serine availability, receptor activation, and downstream signaling cascades. This multilevel analysis approach advances our understanding of how D-serine signaling contributes to synaptic plasticity, learning and memory, and how its dysregulation may contribute to conditions like schizophrenia, epilepsy, and neurodegenerative disorders.

What future directions are emerging for SRR research using fluorescently-labeled antibodies?

The future of SRR research utilizing fluorescently-labeled antibodies like the FITC-conjugated variant is poised for significant advancement through integration with cutting-edge technologies. Super-resolution microscopy techniques—including STED, STORM, and PALM—will enable visualization of SRR localization at the nanoscale level, potentially revealing previously undetectable subcellular compartmentalization or protein-protein interactions that influence D-serine production. The emerging field of spatial transcriptomics combined with multiplex immunofluorescence will allow simultaneous visualization of SRR protein expression and mRNA transcription patterns across entire brain sections, providing insights into transcriptional and post-transcriptional regulation mechanisms. Advanced tissue clearing methods compatible with fluorescent proteins, such as CLARITY, iDISCO, and CUBIC, will facilitate whole-brain imaging of SRR distribution across intact neural circuits, revealing region-specific expression patterns relevant to various cognitive functions. In vivo antibody-based imaging approaches using minimally invasive techniques might eventually allow longitudinal tracking of SRR expression changes during disease progression or following therapeutic interventions. The application of artificial intelligence and machine learning algorithms to analyze complex immunofluorescence datasets will enable identification of subtle patterns in SRR expression that correlate with specific physiological states or pathological conditions. Additionally, the development of photoactivatable or photoswitchable fluorescent conjugates for SRR antibodies could allow selective visualization and tracking of enzyme populations in defined neural circuits. These technological advancements, combined with increasing knowledge of SRR's role in neurological function and dysfunction, will drive new therapeutic approaches targeting the D-serine pathway for conditions ranging from schizophrenia to neurodegenerative diseases and beyond.