serhl Antibody

What is SERHL and its human ortholog?

SERHL is a protein primarily expressed in mice, while its human ortholog is serine hydrolase like 2 (SERHL2). SERHL2 belongs to the AB hydrolase protein superfamily and functions as a suspected serine hydrolase. This protein is encoded by the SERHL2 gene (Gene ID: 253190) in humans and serves as a cellular marker for characterizing astrocytes in neurological research . The human protein has synonyms including dJ222E13.1, serine hydrolase-like protein 2, and testis secretory sperm-binding protein Li 216e .

What is the protein structure and family classification of SERHL?

SERHL2 (Protein ID: Q9H4I8) is classified as a member of the AB hydrolase protein superfamily. The protein functions as a serine hydrolase, containing catalytic domains characteristic of hydrolytic enzymes. Its structure allows it to perform hydrolytic reactions typical of serine proteases, though the specific substrates and comprehensive enzymatic activities remain areas of active investigation . The protein's classification within this family suggests its potential role in various cellular processes including metabolism and signal transduction.

What applications are SERHL antibodies commonly used for?

SERHL antibodies are primarily used in Western Blot (WB) and ELISA applications. According to manufacturer specifications, typical dilution ranges are 1:500-2000 for Western Blot and 1:5000-10000 for ELISA applications . Some antibodies may also be suitable for immunoprecipitation (IP) depending on the specific product . These applications allow researchers to detect and quantify SERHL protein expression in various experimental contexts, particularly for studies involving neural tissues where SERHL serves as an astrocyte marker.

How should SERHL antibodies be validated before experimental use?

For proper validation of SERHL antibodies, researchers should perform a comprehensive characterization process including:

• Specificity testing: Run Western blots with positive and negative control samples to confirm target recognition.

• Cross-reactivity assessment: Test the antibody against related proteins, particularly other serine hydrolases.

• Application-specific validation: For each intended application (WB, ELISA, IP), perform validation using appropriate controls.

• Sensitivity determination: Establish detection limits using serial dilutions of target protein.

• Reproducibility testing: Confirm consistent performance across multiple experiments.

This validation approach helps ensure experimental reliability and prevents misleading results due to non-specific binding .

What are the optimal storage and handling conditions for SERHL antibodies?

SERHL antibodies should typically be stored at -20°C for long-term preservation of activity . For working solutions, aliquoting is recommended to avoid repeated freeze-thaw cycles which can degrade antibody performance. When handling, maintain sterile conditions and avoid contamination. Most commercial SERHL antibodies are formulated as liquids with stabilizers to maintain integrity, with typical concentrations around 1 mg/ml . Prior to use, allow the antibody to equilibrate to room temperature and gently mix without vortexing to prevent protein denaturation.

What controls should be included when using SERHL antibodies for Western blot?

When designing Western blot experiments with SERHL antibodies, include the following controls:

Including these controls helps differentiate between specific signal and experimental artifacts, particularly important when working with polyclonal SERHL antibodies that may have varied epitope recognition .

How do antibody titers correlate with detection sensitivity in SERHL research?

Antibody titers significantly impact detection sensitivity in SERHL research. Higher antibody concentrations generally improve detection limits but may increase background signal. In neutralization studies with other antibodies, endpoint titers of approximately 1:40 in live virus microneutralization assays have been shown to correspond with detection thresholds in lateral flow immunoassays . For SERHL antibodies specifically, dilution studies have demonstrated that Western blot applications typically require higher antibody concentrations (1:500-2000) compared to ELISA applications (1:5000-10000) . This differential in optimal concentration reflects the varying detection sensitivities across experimental platforms and should be considered when designing experimental protocols.

What are the challenges in cross-species reactivity when using SERHL antibodies?

Cross-species reactivity presents significant challenges in SERHL antibody applications. Available commercial antibodies show varied reactivity profiles across species: some are specific to human SERHL2, while others demonstrate reactivity with zebrafish or other fish models . This variability stems from evolutionary differences in the SERHL protein sequence across species. When conducting comparative studies across different model organisms, researchers should:

• Specifically select antibodies validated for cross-reactivity with target species

• Perform preliminary validation experiments to confirm reactivity in each species

• Consider epitope mapping to identify conserved regions across species

• Adjust experimental protocols (blocking conditions, antibody concentrations) to optimize for cross-species applications

• Include appropriate species-specific positive controls

These considerations are particularly important when translating findings between mouse models and human applications .

How can researchers distinguish between SERHL isoforms using antibody-based approaches?

Distinguishing between SERHL isoforms requires careful selection of antibodies targeting isoform-specific epitopes. Researchers should:

• Select antibodies raised against unique regions of specific isoforms rather than conserved domains

• Employ immunoprecipitation followed by mass spectrometry to identify exact isoforms present

• Conduct Western blot analysis with high-resolution gels to separate closely related isoforms by molecular weight

• Perform pre-adsorption experiments with recombinant isoforms to determine antibody specificity

• Consider complementary approaches such as RT-PCR to identify isoform-specific transcripts

This multi-faceted approach allows for accurate identification and quantification of specific SERHL isoforms, which is crucial when investigating their distinct functions in different cellular contexts .

What are common sources of false positives/negatives when using SERHL antibodies?

Common sources of false results when working with SERHL antibodies include:

False Positives:

• Cross-reactivity with other serine hydrolases due to structural similarities

• Non-specific binding to highly abundant proteins

• Excessive antibody concentration leading to background signal

• Sample contamination with endogenous phosphatases or peroxidases

• Insufficient blocking or washing steps in immunoassays

False Negatives:

• Protein denaturation affecting epitope accessibility

• Insufficient antigen retrieval in fixed samples

• Antibody degradation due to improper storage

• Target protein expression below detection threshold

• Interference from sample buffer components

To minimize these issues, researchers should optimize protocols for each specific application and include appropriate controls to distinguish between true and false signals .

How can batch-to-batch variability in SERHL antibodies be addressed?

Batch-to-batch variability is a significant concern with antibody reagents, particularly polyclonal antibodies like many SERHL antibodies . To address this challenge:

• Maintain detailed records of antibody lot numbers and performance characteristics

• Perform validation tests when switching to a new lot

• Create internal reference standards from well-characterized lots

• Consider purchasing larger quantities of a single lot for long-term studies

• Implement normalization procedures using consistent control samples

• When possible, use recombinant monoclonal antibodies which offer greater consistency

• Communicate with manufacturers about observed variability

Additionally, researchers should consider developing quantitative metrics for antibody performance to objectively assess and compare different batches .

What are the critical differences between polyclonal and monoclonal SERHL antibodies in research applications?

Polyclonal and monoclonal SERHL antibodies offer distinct advantages and limitations for research:

Currently, polyclonal SERHL antibodies are more widely available commercially . The choice between polyclonal and monoclonal depends on research needs—polyclonals offer robust detection across applications while monoclonals provide greater specificity for discriminating between closely related proteins or specific epitopes .

How can SERHL antibodies be effectively used in multiplex immunoassays?

Implementing SERHL antibodies in multiplex immunoassays requires careful optimization to maintain specificity while enabling simultaneous detection of multiple targets. Researchers should:

• Select SERHL antibodies with minimal cross-reactivity to other targets in the multiplex panel

• Perform preliminary singleplex assays to establish baseline performance metrics

• Titrate antibody concentrations to achieve balanced signal intensity across all targets

• Use antibodies from different host species when possible to enable species-specific secondary detection

• Employ spectral unmixing for fluorescently labeled antibodies to minimize signal overlap

• Validate multiplex results against traditional singleplex assays

These approaches enable researchers to simultaneously examine SERHL alongside other biomarkers, particularly valuable when investigating its role in complex biological processes like astrocyte function .

What are the emerging applications of SERHL antibodies in neurological research?

SERHL antibodies are increasingly valuable in neurological research applications due to SERHL's role as an astrocyte marker . Emerging applications include:

• Single-cell proteomic analysis of neural cell populations

• Spatial transcriptomic studies correlating SERHL expression with location in neural tissues

• Investigation of astrocyte heterogeneity in various neurological conditions

• Tracking astrocyte activation in response to neuroinflammatory triggers

• Development of astrocyte-specific isolation methods using SERHL-targeted approaches

• Screening potential therapeutic compounds that modulate astrocyte function

These applications are expanding our understanding of astrocyte biology and potentially identifying new therapeutic targets for neurological disorders where astrocyte dysfunction plays a role .

How does epitope selection influence the performance of SERHL antibodies in different applications?

Epitope selection critically influences SERHL antibody performance across applications. The choice of target epitope affects:

• Accessibility in different applications: Epitopes located in structured domains may be accessible in denatured states (Western blot) but hidden in native conformations (immunoprecipitation).

• Cross-reactivity profile: Targeting highly conserved regions increases cross-species reactivity but may reduce specificity within the serine hydrolase family.

• Functional interference: Antibodies targeting catalytic domains may inhibit enzymatic activity, which could be advantageous for functional studies but problematic for detecting active enzyme.

• Post-translational modification sensitivity: Epitopes containing phosphorylation or glycosylation sites may be inaccessible when these modifications are present.

• Stability in fixation procedures: Some epitopes may be more resistant to chemical fixatives used in immunohistochemistry.

Researchers should select antibodies with epitopes appropriate for their specific application, considering both the structural state of the protein and the experimental conditions .