DHRS12 Antibody

What is DHRS12 and why is it studied?

DHRS12 (Dehydrogenase/Reductase SDR Family Member 12) is a member of the short-chain dehydrogenase/reductase (SDR) family, also known as SDR40C1 . It functions as an enzyme involved in redox reactions and is classified under the Enzyme Commission Number EC:1.1.-.- . DHRS12 is studied primarily for its potential roles in cellular metabolism and redox homeostasis within human tissues . The gene is identified by GeneID 79758 and is mapped to the human genome with UniProt Primary Accession Number A0PJE2 . Research on DHRS12 contributes to our understanding of dehydrogenase/reductase activity in normal cellular function and potential disease states.

What types of DHRS12 antibodies are available for research?

Based on current research materials, the primary type of DHRS12 antibody available is polyclonal, specifically rabbit polyclonal antibodies that target human DHRS12 . These antibodies are available in both unconjugated forms and conjugated versions (such as HRP-conjugated) . The immunogen used for developing these antibodies typically consists of recombinant human DHRS12 protein spanning amino acids 1-271 . The antibodies are predominantly presented in liquid form with high purity (>95%) and are purified using protein G or affinity purification methods .

What are the common applications for DHRS12 antibodies?

DHRS12 antibodies are validated for multiple research applications with varying recommended dilutions:

These applications allow researchers to investigate DHRS12 expression patterns, cellular localization, and potential interactions with other proteins . The antibodies have been specifically validated for human samples, making them suitable for studies using human cell lines and tissues .

How should DHRS12 antibodies be stored and handled?

Proper storage and handling are critical for maintaining antibody functionality. DHRS12 antibodies should be aliquoted upon receipt and stored at -20°C to avoid repeated freeze-thaw cycles that can degrade antibody quality . For HRP-conjugated antibodies, it's particularly important to avoid exposure to light during storage . The antibodies are typically supplied in a buffer containing 0.01M PBS (pH 7.4), 0.03% Proclin-300, and 50% glycerol . When working with these antibodies, researchers should follow standard laboratory practices for handling antibody solutions, including using appropriate personal protective equipment and maintaining sterile conditions when possible .

What are the optimal validation controls for DHRS12 antibody experiments?

For rigorous validation of DHRS12 antibody experiments, researchers should implement multiple control strategies. Positive controls should include samples known to express DHRS12, such as relevant human cell lines or tissue samples where expression has been previously documented . Negative controls should include samples where DHRS12 expression is expected to be absent or samples treated with non-specific IgG from the same host species (rabbit) . For knockdown validation, researchers should consider using DHRS12 siRNA or CRISPR-mediated knockouts to demonstrate antibody specificity .

Some DHRS12 antibodies have undergone specificity verification using protein arrays containing the target protein plus 383 other non-specific proteins, which provides strong evidence for antibody specificity . For immunocytochemistry experiments, researchers should consider using competitive blocking with the immunizing peptide to further validate specificity . These validation approaches ensure reliable and reproducible results when working with DHRS12 antibodies.

How do different fixation methods affect DHRS12 antibody performance in immunostaining applications?

Fixation methodology significantly impacts DHRS12 antibody performance in immunocytochemistry and immunohistochemistry applications. While the search results don't provide specific fixation recommendations for DHRS12 antibodies, general principles apply based on antibody characteristics. For polyclonal DHRS12 antibodies targeting human samples, researchers should consider testing both cross-linking fixatives (like paraformaldehyde) and precipitating fixatives (like methanol/acetone) .

When optimizing fixation protocols, researchers should evaluate factors such as epitope accessibility, signal-to-noise ratio, and cellular morphology preservation . Since the available DHRS12 antibodies are developed against recombinant proteins spanning amino acids 1-271, they likely recognize multiple epitopes, some of which may be more sensitive to particular fixation methods . For immunofluorescence applications, the recommended dilution range of 1:50 - 1:200 provides a starting point, but researchers should conduct titration experiments with different fixation methods to determine optimal conditions for their specific experimental system .

What factors should be considered when troubleshooting non-specific binding of DHRS12 antibodies?

When encountering non-specific binding with DHRS12 antibodies, researchers should systematically evaluate several factors. First, antibody concentration is critical – the recommended dilutions (WB: 1:500-1:2000, IHC: 1:20-1:200, IF: 1:50-1:200) provide starting points, but optimal concentrations may vary based on sample type and experimental conditions . Higher antibody concentrations often increase non-specific binding.

Blocking protocols significantly impact specificity; researchers should optimize blocking agent composition (BSA, normal serum, commercial blockers) and blocking time . The buffer composition used for antibody dilution can also affect specificity; the antibodies are supplied in PBS with Proclin-300 and glycerol, but different diluents may be beneficial for reducing background in specific applications .

Cross-reactivity potential should be considered, though the available DHRS12 antibodies have been primarily validated against human samples . For tissue sections or cell samples with high endogenous peroxidase or phosphatase activity, appropriate quenching steps should be implemented before antibody application . Additional washing steps with optimized washing buffers can also help reduce non-specific binding without compromising specific signal detection .

How can DHRS12 antibodies be utilized in multiplexed immunoassays with other biomarkers?

DHRS12 antibodies can be incorporated into multiplexed immunoassays through careful planning and optimization. For immunofluorescence multiplexing, researchers should select DHRS12 antibodies with compatible host species or isotypes relative to other target antibodies . The unconjugated rabbit polyclonal DHRS12 antibodies can be paired with mouse monoclonal antibodies against other targets, followed by species-specific secondary antibodies with different fluorophores .

HRP-conjugated DHRS12 antibodies offer direct detection capabilities that can be integrated into sequential immunoenzyme detection schemes . For these approaches, researchers should optimize antibody stripping or inactivation protocols between detection steps to prevent cross-reactivity . When designing multiplexed experiments, researchers should verify that the subcellular localization pattern of DHRS12 is distinct from other biomarkers to facilitate clear interpretation of results .

Cross-validation of results using complementary techniques such as Western blotting alongside immunofluorescence provides stronger evidence for co-expression or co-localization patterns observed in multiplexed assays . This multi-technique approach helps control for potential artifacts that might arise in any single detection method.

What protocol modifications are recommended for detecting low-abundance DHRS12 in different sample types?

For detecting low-abundance DHRS12, researchers should implement several protocol modifications. For Western blot applications, increasing protein loading (up to 50-100 μg per lane), using enhanced chemiluminescence detection systems, and extending primary antibody incubation times (overnight at 4°C) can improve sensitivity . The recommended antibody dilution of 1:500 represents the highest concentration within the suggested range (1:500-1:2000) and would be appropriate for low-abundance detection .

For immunohistochemistry and immunofluorescence, signal amplification systems such as tyramide signal amplification or fluorescent-labeled secondary antibody complexes can significantly enhance detection sensitivity . Heat-induced epitope retrieval methods should be optimized for tissue sections, with citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) as starting conditions . For DHRS12 detection in tissues with high background, biotin-free detection systems are recommended to avoid endogenous biotin interference .

When working with challenging sample types, researchers might consider concentrating the sample through immunoprecipitation prior to Western blot analysis or using more sensitive detection methods like proximity ligation assay for in situ detection of low-abundance DHRS12 .

How can researchers reliably quantify DHRS12 expression using antibody-based methods?

Reliable quantification of DHRS12 expression requires standardized approaches across multiple techniques. For Western blot quantification, researchers should use internal loading controls (like GAPDH, β-actin) and implement standard curves using recombinant DHRS12 protein at known concentrations . Densitometric analysis should be performed using specialized software with appropriate background subtraction .

For immunohistochemical quantification, researchers should establish standardized scoring systems based on staining intensity and percentage of positive cells . Digital image analysis using specialized software can provide more objective quantification than manual scoring . For immunofluorescence quantification, fluorescence intensity measurements should be performed under standardized acquisition parameters across all samples .

ELISA-based quantification offers the most direct approach for DHRS12 protein quantification, with available DHRS12 antibodies validated for ELISA applications . Researchers should develop standard curves using recombinant DHRS12 protein and implement technical replicates (minimum triplicates) to ensure statistical reliability . Regardless of the quantification method, researchers should validate findings across multiple antibody-based techniques when possible, as each method has inherent limitations and potential artifacts .

What technical considerations are important when selecting secondary antibodies for DHRS12 detection?

When selecting secondary antibodies for DHRS12 detection, researchers should consider several technical aspects. Since the available DHRS12 antibodies are rabbit polyclonal IgG, secondary antibodies should specifically target rabbit IgG with minimal cross-reactivity to other species . For immunofluorescence applications, fluorophore selection should align with the available microscopy setup, avoiding spectral overlap with other fluorescent markers in multiplexed experiments .

For Western blot applications, HRP-conjugated anti-rabbit secondaries are commonly used, but alkaline phosphatase conjugates can offer advantages for certain detection systems . Secondary antibody concentration requires careful optimization – too high concentrations increase background, while too low concentrations reduce specific signal . A starting dilution range of 1:1000-1:5000 is typical, but should be adjusted based on specific experimental conditions and primary antibody concentration .

Pre-adsorbed secondary antibodies are recommended when working with samples containing proteins from multiple species to minimize cross-reactivity . For tissues with high endogenous biotin or when using biotin-based detection systems, biotin blocking steps should be implemented to reduce background . Secondary antibody storage and handling also impact performance – aliquoting to avoid freeze-thaw cycles and protecting fluorophore-conjugated antibodies from light exposure are essential practices .

How do DHRS12 expression patterns compare across different human tissues and cell types?

While the search results don't provide specific data on DHRS12 expression patterns across tissues, researchers investigating this question should use validated DHRS12 antibodies in systematic tissue profiling studies . Based on general characteristics of SDR family members, DHRS12 expression may vary significantly across tissue types with potential enrichment in metabolically active tissues .

For comparative expression analysis, researchers should implement standardized protocols across all samples, using the same antibody lot, concentration, and detection system . Quantitative approaches like tissue microarrays with the DHRS12 antibody (dilution 1:20-1:200) would allow efficient screening across multiple tissue types . Complementary approaches such as Western blotting of tissue lysates and qPCR analysis provide validation across methodologies .

When interpreting expression patterns, researchers should consider that post-translational modifications might affect antibody recognition, potentially resulting in tissue-specific variations in detection efficiency . Therefore, multiple antibody clones or epitopes should ideally be tested when possible to ensure comprehensive detection .

What are the best practices for correlating DHRS12 protein expression with functional outcomes?

To establish meaningful correlations between DHRS12 expression and functional outcomes, researchers should implement multi-faceted approaches. Antibody-based detection of DHRS12 protein expression should be paired with functional assays that measure dehydrogenase/reductase activity, as DHRS12 belongs to the SDR enzyme family (EC:1.1.-.-) .

Gain-of-function and loss-of-function experimental designs offer powerful approaches for establishing causal relationships . After manipulating DHRS12 expression levels, researchers can use the validated antibodies (WB: 1:500-1:2000) to confirm expression changes, then measure functional parameters . For loss-of-function studies, siRNA or CRISPR-based approaches followed by antibody validation of knockdown efficiency provide robust experimental designs .

Co-immunoprecipitation using DHRS12 antibodies can identify protein interaction partners that might mediate functional effects . For these experiments, researchers should optimize antibody concentration and binding conditions to ensure efficient immunoprecipitation without disrupting physiologically relevant interactions . When interpreting correlative data, researchers should consider potential confounding factors such as compensatory expression of other SDR family members and context-dependent effects that might influence functional outcomes .

How can researchers address data discrepancies when using different DHRS12 antibody clones or suppliers?

When encountering discrepancies between different DHRS12 antibody sources, researchers should implement systematic validation approaches. First, detailed comparison of antibody specifications is essential – examining immunogen sequences, host species, and clonality across different sources . The available DHRS12 antibodies use similar immunogens (recombinant human DHRS12 protein, amino acids 1-271), but even slight variations in epitope regions can affect recognition patterns .

Side-by-side validation experiments are crucial, testing multiple antibodies under identical conditions across several applications . These should include positive controls (samples known to express DHRS12) and negative controls (samples with DHRS12 knockdown) . Western blot analysis can reveal potential differences in specificity based on band patterns and molecular weights .

Epitope mapping studies, while more resource-intensive, can definitively resolve discrepancies by identifying exactly which regions of DHRS12 are recognized by different antibodies . Researchers encountering persistent discrepancies should consider using complementary detection methods like mass spectrometry to provide antibody-independent validation .

When reporting results from DHRS12 studies, researchers should explicitly state which antibody was used (including catalog number and lot number if possible) to facilitate comparison across studies and improve reproducibility .

What emerging technologies are enhancing DHRS12 antibody applications in research?

Several emerging technologies are expanding the utility of DHRS12 antibodies in research applications. Advanced imaging techniques like super-resolution microscopy and expansion microscopy are enabling more detailed analysis of DHRS12 subcellular localization beyond the capabilities of standard immunofluorescence approaches (typically used at 1:50-1:200 dilution) . These techniques require careful optimization of antibody concentration and fixation protocols to maintain specificity while improving resolution .

Single-cell proteomics approaches that incorporate antibody-based detection are allowing researchers to examine DHRS12 expression heterogeneity within seemingly homogeneous cell populations . Proximity labeling techniques using DHRS12 antibodies can reveal spatial interaction networks that provide functional insights beyond traditional co-immunoprecipitation approaches .

Microfluidic antibody-based assays are improving sensitivity and reducing sample requirements for DHRS12 detection, potentially allowing analysis from limited clinical specimens . As these technologies continue to evolve, researchers should validate each new application with appropriate controls to ensure that the fundamental specificity of DHRS12 antibodies is maintained across these innovative platforms .

What considerations are important when designing longitudinal studies monitoring DHRS12 expression?

For longitudinal studies monitoring DHRS12 expression, researchers should implement several key strategies to ensure data consistency. Antibody lot consistency is paramount – researchers should secure sufficient quantities of a single lot for the entire study duration or perform cross-lot validation if multiple lots must be used . Sample collection, processing, and storage protocols should be standardized to minimize technical variables that could confound biological changes in DHRS12 expression .

Reference standards should be included in each experimental batch – these could include pooled reference samples or recombinant DHRS12 protein standards that allow normalization across experiments . When analyzing Western blot data longitudinally, consistent loading controls and standardized quantification approaches are essential . For immunohistochemistry or immunofluorescence in longitudinal studies, automated staining platforms can reduce batch effects, and digital image analysis using consistent parameters improves quantification reliability .