C19orf44 Antibody

What is C19orf44 and what are its key characteristics?

C19orf44 (Chromosome 19 Open Reading Frame 44) is an uncharacterized protein encoded by a gene located on chromosome 19. The protein has a calculated molecular weight of approximately 66 kDa, though the observed molecular weight in Western blot analyses typically ranges from 66-71 kDa . It is expressed in various tissues including human cell lines (such as L02 cells), mouse liver tissue, and rat brain tissue . The protein's full sequence is available under GenBank Accession Number BC027869 . Although its specific function remains largely undetermined, it represents an area of active research interest, particularly in comparative biology given its conservation across multiple species.

What applications are most suitable for C19orf44 antibodies?

C19orf44 antibodies have been validated for multiple research applications with varying recommended dilutions:

When selecting an application, researchers should consider that WB applications have shown consistent results across multiple species, making this technique particularly reliable for C19orf44 detection . For optimal results, it is recommended to conduct preliminary experiments to determine the ideal antibody concentration for your specific experimental conditions and sample types.

What species reactivity is confirmed for C19orf44 antibodies?

Most commercial C19orf44 antibodies demonstrate confirmed reactivity with human, mouse, and rat samples . Positive Western blot detection has been specifically validated in:

• Human L02 cells

• Mouse liver tissue

• Rat brain tissue

Researchers working with other species should be aware that cross-reactivity has not been comprehensively tested across all mammalian models. When studying C19orf44 in non-validated species, preliminary validation experiments are strongly recommended before proceeding with full experimental workflows.

How should C19orf44 antibodies be properly stored and handled?

Proper storage and handling of C19orf44 antibodies is critical for maintaining reactivity and specificity:

• Store at -20°C for long-term preservation

• Most preparations include PBS with 0.02% sodium azide and 50% glycerol at pH 7.3

• Antibodies remain stable for approximately one year after shipment when stored under recommended conditions

• Important note: Some manufacturers specify "DO NOT ALIQUOT" for certain preparations , while others indicate that aliquoting is unnecessary for -20°C storage

• Some formulations may contain 0.1% BSA in smaller size formats (e.g., 20μl)

When working with these antibodies, avoid repeated freeze-thaw cycles, minimize exposure to light, and handle according to standard laboratory safety procedures, particularly considering the presence of sodium azide in the storage buffer.

What are the optimal Western blot protocols for C19orf44 antibody applications?

For optimal Western blot results with C19orf44 antibodies, researchers should follow these methodological guidelines:

• Sample preparation:

  • Lyse cells/tissues in RIPA buffer containing protease inhibitors

  • Determine protein concentration using BCA or Bradford assay

  • Load 20-40 μg of total protein per lane

• Gel electrophoresis and transfer:

  • Use 10% SDS-PAGE gels for optimal resolution around the 66-71 kDa range

  • Transfer to PVDF membrane (preferred over nitrocellulose for this protein)

• Antibody incubation:

  • Block membrane with 5% non-fat milk in TBST for 1 hour at room temperature

  • Incubate with primary C19orf44 antibody at dilutions of 1:500-1:1000 overnight at 4°C

  • Wash 3-5 times with TBST, 5 minutes each

  • Incubate with HRP-conjugated secondary antibody at 1:5000-1:10000 for 1 hour at room temperature

• Detection:

  • Develop using enhanced chemiluminescence (ECL) substrate

  • Expected band size: 66-71 kDa

This protocol has been validated using L02 cells, mouse liver tissue, and rat brain tissue as positive controls . When troubleshooting, remember that the observed molecular weight (66-71 kDa) may vary slightly due to post-translational modifications or splice variants.

How can antibody specificity for C19orf44 be validated before use in critical experiments?

Thorough validation of C19orf44 antibody specificity is essential before conducting critical experiments. A comprehensive validation approach should include:

• Positive and negative control samples:

  • Use known positive controls such as L02 cells, mouse liver tissue, or rat brain tissue

  • Include negative controls (tissues/cells with low or no C19orf44 expression)

  • Consider siRNA/shRNA knockdown controls to confirm specificity

• Multiple detection methods:

  • Compare results across different applications (WB, IHC, ICC-IF) when possible

  • Use at least two different antibodies targeting distinct epitopes of C19orf44

• Molecular weight verification:

  • Confirm single band at the expected 66-71 kDa range

  • Be aware that post-translational modifications may affect migration pattern

• Peptide competition assays:

  • Pre-incubate antibody with immunizing peptide/protein (such as the recombinant protein with Accession Number BC027869)

  • Verify elimination or significant reduction of signal

• Sequence alignment analysis:

  • Perform in silico analysis to ensure epitope uniqueness

  • Check for potential cross-reactivity with homologous proteins

For optimal validation, researchers should document all validation steps and include appropriate controls in each experiment. This approach significantly enhances the reliability and reproducibility of research findings involving C19orf44.

What factors influence the performance of C19orf44 antibodies in immunohistochemistry?

Multiple factors can significantly impact the performance of C19orf44 antibodies in immunohistochemistry (IHC) applications:

• Fixation methods:

  • Formalin fixation duration affects epitope accessibility

  • Overfixation may mask epitopes and reduce signal intensity

  • Different epitopes show varying sensitivity to fixation conditions

• Antigen retrieval techniques:

  • Heat-induced epitope retrieval (HIER) using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0)

  • Enzymatic retrieval methods may be necessary for certain tissues

  • Optimization of retrieval conditions is often tissue-specific

• Antibody dilution:

  • Recommended starting dilution for IHC is 1:200-1:500

  • Titration experiments are essential for each new batch of antibody

  • Different tissue types may require different antibody concentrations

• Detection systems:

  • Polymer-based detection systems often provide superior signal-to-noise ratio

  • Tyramide signal amplification may be beneficial for low abundance targets

  • Chromogenic vs. fluorescent detection considerations

• Tissue-specific factors:

  • Endogenous peroxidase or phosphatase activity can cause background

  • Autofluorescence in certain tissues requires appropriate quenching

  • Tissue-specific blocking reagents may be necessary

For consistent and reliable IHC results with C19orf44 antibodies, researchers should systematically optimize each of these parameters and maintain rigorous documentation of protocol conditions. This methodological approach enables meaningful comparison across different experiments and research groups.

How can researchers troubleshoot inconsistent results with C19orf44 antibodies?

When encountering inconsistent results with C19orf44 antibodies, implement this systematic troubleshooting approach:

• Antibody-related issues:

  • Verify antibody storage conditions (maintain at -20°C, avoid repeated freeze-thaw cycles)

  • Check antibody expiration date and lot number

  • Consider testing a new antibody lot or alternative clone

  • Verify compatibility with host species of secondary antibody

• Sample preparation concerns:

  • Ensure complete protein denaturation for WB applications

  • Verify protein concentration estimation methods

  • Check sample degradation by probing for housekeeping proteins

  • Ensure proper tissue fixation for IHC/ICC applications

• Protocol optimization:

  • Adjust antibody dilution (try a concentration series from 1:200 to 1:2000)

  • Modify incubation time and temperature

  • Test different blocking reagents to reduce background

  • Optimize antigen retrieval methods for IHC

• Detection system evaluation:

  • Compare different secondary antibodies

  • Assess alternative detection chemistries

  • Consider signal amplification systems for weak signals

• Analytical considerations:

  • Document any variation in band pattern (potential splice variants)

  • Compare with published literature on C19orf44 detection

  • Consider post-translational modifications affecting detection

Maintaining detailed records of all experimental conditions facilitates efficient troubleshooting and protocol optimization when working with C19orf44 antibodies.

What experimental controls are essential when studying C19orf44 expression patterns?

Robust experimental design for C19orf44 expression studies requires inclusion of these critical controls:

• Positive tissue/cell controls:

  • L02 cells, mouse liver tissue, and rat brain tissue have demonstrated reliable C19orf44 expression

  • Include these validated positive controls alongside experimental samples

• Negative controls:

  • Primary antibody omission controls

  • Isotype controls (normal rabbit IgG at equivalent concentration)

  • Tissues/cells known or expected to have minimal C19orf44 expression

• Loading/procedural controls:

  • Housekeeping proteins (β-actin, GAPDH, tubulin) for Western blot normalization

  • Internal tissue controls for IHC (tissues with known expression patterns)

  • Procedural controls to validate staining/detection protocols

• Validation controls:

  • siRNA/shRNA knockdown of C19orf44 to demonstrate specificity

  • Recombinant protein competition assays using the immunizing antigen (BC027869)

  • Secondary antibody-only controls to assess non-specific binding

• Cross-validation approaches:

  • Comparison of results across multiple detection methods (WB, IHC, ICC)

  • Use of alternative antibodies targeting different epitopes of C19orf44

  • Correlation with mRNA expression data when available

How should researchers approach comparative analysis of C19orf44 across different species?

When conducting comparative studies of C19orf44 across species, researchers should implement the following methodological approach:

• Antibody cross-reactivity validation:

  • Verify that the selected antibody has confirmed reactivity with target species

  • Current literature confirms reactivity with human, mouse, and rat samples

  • For other species, conduct preliminary validation experiments

• Sequence homology considerations:

  • Analyze the sequence conservation of C19orf44 across target species

  • Identify conserved and variable regions that might affect antibody binding

  • Select antibodies targeting highly conserved epitopes for cross-species studies

• Application-specific adaptations:

  • Adjust protein extraction methods based on tissue-specific characteristics

  • Modify blocking reagents to address species-specific background issues

  • Adapt antigen retrieval methods for species-specific tissue architecture

• Normalization strategies:

  • Use species-appropriate housekeeping genes/proteins for normalization

  • Consider species-specific expression levels of reference genes

  • Implement statistical methods that account for inter-species variability

• Interpretation guidelines:

  • Consider species-specific post-translational modifications

  • Account for potential differences in subcellular localization

  • Interpret apparent molecular weight variations (66-71 kDa range) in evolutionary context

By implementing these methodological considerations, researchers can generate more reliable comparative data on C19orf44 expression and function across different species, contributing to broader understanding of its evolutionary conservation and potential biological roles.

What are the key considerations for quantitative analysis of C19orf44 expression?

For rigorous quantitative analysis of C19orf44 expression, researchers should address the following methodological considerations:

• Western blot quantification:

  • Use a dilution series to establish antibody linearity range

  • Implement optimal blocking conditions to minimize background

  • Employ software-based densitometry with appropriate background subtraction

  • Normalize to validated housekeeping proteins

  • Report relative expression values rather than absolute densitometric units

• Immunohistochemistry quantification:

  • Define specific scoring parameters (intensity, percentage positive cells)

  • Use digital image analysis software for unbiased assessment

  • Implement double-blind scoring when manual quantification is necessary

  • Establish clear cutoff values for positivity

  • Report both distribution patterns and intensity metrics

• ELISA-based quantification:

  • Generate standard curves using recombinant C19orf44 protein

  • Determine assay sensitivity and dynamic range

  • Validate assay specificity using knockout/knockdown samples

  • Perform technical and biological replicates

  • Calculate intra- and inter-assay coefficients of variation

• Statistical analysis considerations:

  • Select appropriate statistical tests based on data distribution

  • Account for multiple comparisons when analyzing expression across tissues

  • Report effect sizes alongside p-values

  • Consider hierarchical or mixed models when analyzing nested data

  • Implement power analysis to determine adequate sample sizes

• Reporting standards:

  • Document all antibody details (catalog number, lot, dilution)

  • Specify image acquisition parameters

  • Include representative images of high, medium, and low expression

  • Present raw data alongside processed/normalized results when possible

Adherence to these quantitative guidelines ensures generation of reproducible and statistically sound data on C19orf44 expression across experimental conditions.

How can C19orf44 antibodies be utilized in protein-protein interaction studies?

C19orf44 antibodies can be valuable tools for investigating protein-protein interactions through these methodological approaches:

• Co-immunoprecipitation (Co-IP):

  • Use C19orf44 antibodies for immunoprecipitation of native protein complexes

  • Starting dilution recommendation: 1:50-1:200 of antibody solution

  • Use gentle lysis buffers to preserve protein-protein interactions

  • Confirm antibody suitability for IP applications before proceeding

  • Include IgG control immunoprecipitations to identify non-specific interactions

• Proximity ligation assay (PLA):

  • Combine C19orf44 antibody with antibodies against suspected interacting partners

  • Requires antibodies raised in different host species

  • Provides subcellular localization information for interaction events

  • Optimization of antibody dilutions is critical (start with 1:200-1:500)

  • Include appropriate negative controls (single antibody, non-interacting protein pairs)

• Immunofluorescence co-localization:

  • Use C19orf44 antibodies in combination with antibodies against potential interactors

  • Apply appropriate spectral unmixing for multi-color imaging

  • Implement quantitative co-localization analysis (Pearson's coefficient, Manders' overlap)

  • Control for random co-localization through spatial randomization tests

  • Consider super-resolution microscopy for detailed interaction studies

• Protein complex analysis:

  • Apply C19orf44 antibodies for Western blot analysis of native protein complexes

  • Consider blue native PAGE or gradient gel approaches

  • Use chemical crosslinking to stabilize transient interactions

  • Combine with mass spectrometry for unbiased interactome analysis

  • Validate key interactions through reciprocal co-immunoprecipitation

These methodological approaches enable researchers to investigate the functional protein interaction network of C19orf44, potentially revealing insights into its biological roles and mechanisms of action.

What considerations should be made when designing immunohistochemical studies for tissue distribution of C19orf44?

When designing immunohistochemical studies to map C19orf44 tissue distribution, researchers should consider these methodological aspects:

• Tissue selection and preparation:

  • Include diverse tissue types for comprehensive distribution profiling

  • Consider developmental stages for temporal expression analysis

  • Use standardized fixation protocols (10% neutral buffered formalin is recommended)

  • Optimize fixation time to preserve antigenicity while maintaining morphology

  • Process tissue microarrays (TMAs) for high-throughput screening across multiple tissues

• Antibody selection and validation:

  • Use antibodies validated specifically for IHC applications (e.g., HPA041683, dilution 1:200-1:500)

  • Validate antibody specificity using positive controls (L02 cells, liver tissue)

  • Include peptide competition controls to confirm specificity

  • Consider using multiple antibodies targeting different epitopes for cross-validation

• Staining protocol optimization:

  • Systematically compare different antigen retrieval methods

  • Test both chromogenic and fluorescent detection systems

  • Incorporate double-staining with cell type-specific markers

  • Use automated staining platforms when possible to enhance reproducibility

  • Implement stringent washing procedures to minimize background

• Data collection and analysis:

  • Establish a standardized scoring system before beginning analysis

  • Capture high-resolution images of representative areas

  • Document both intensity and subcellular localization patterns

  • Use digital pathology tools for quantitative assessment

  • Correlate IHC findings with available transcriptomic data

• Comprehensive reporting:

  • Document complete methodological details for reproducibility

  • Present findings in the context of normal versus pathological tissues

  • Include detailed descriptions of subcellular localization

  • Report both positive and negative findings across tissue types

  • Provide access to representative images for all tissues examined

By implementing these methodological considerations, researchers can generate comprehensive and reliable data on the tissue distribution patterns of C19orf44, contributing to better understanding of its physiological roles.

How can researchers effectively interpret contradictory results from different C19orf44 antibodies?

When confronted with contradictory results from different C19orf44 antibodies, researchers should implement this systematic analytical approach:

• Antibody characteristics analysis:

  • Compare epitope regions targeted by each antibody

  • Review immunogen sequences (e.g., BC027869)

  • Assess antibody formats (polyclonal vs. monoclonal)

  • Consider host species and purification methods

  • Evaluate validation data provided by manufacturers

• Methodological comparison:

  • Document exact protocols used with each antibody

  • Analyze differences in sample preparation, dilutions, and detection methods

  • Consider application-specific variations (WB vs. IHC optimization)

  • Review incubation conditions and blocking reagents

  • Identify potential procedural sources of discrepancy

• Biological interpretation framework:

  • Consider potential detection of different splice variants

  • Evaluate post-translational modifications affecting epitope accessibility

  • Assess potential protein-protein interactions masking epitopes

  • Review tissue-specific expression patterns and protein conformation

  • Analyze subcellular localization differences

• Validation experiments:

  • Test both antibodies on identical samples simultaneously

  • Implement genetic approaches (siRNA, CRISPR) for specificity confirmation

  • Use recombinant protein with known sequence as a control

  • Perform peptide competition assays with immunizing antigens

  • Consider mass spectrometry validation of detected bands

• Literature integration:

  • Compare findings with published literature on C19orf44

  • Consult antibody validation repositories and databases

  • Reach out to authors of key publications for technical insights

  • Consider collaborative cross-laboratory validation

What are the recommended sample preparation methods for optimal C19orf44 detection?

Optimizing sample preparation is critical for reliable C19orf44 detection across different applications:

• Cell/tissue lysis for Western blot:

  • Use RIPA buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS)

  • Include fresh protease inhibitor cocktail to prevent degradation

  • Perform mechanical disruption (sonication or homogenization) to ensure complete lysis

  • Clarify lysates by centrifugation (14,000 × g, 15 minutes, 4°C)

  • Determine protein concentration using BCA or Bradford assay

  • Store aliquoted samples at -80°C to avoid freeze-thaw cycles

• Tissue preparation for immunohistochemistry:

  • Fix tissues in 10% neutral buffered formalin for 24-48 hours

  • Avoid overfixation which can mask epitopes

  • Process and embed in paraffin following standard protocols

  • Section at 4-5 μm thickness for optimal antibody penetration

  • Use positively charged slides to prevent tissue detachment

  • Include known positive control tissues (liver) in the same block when possible

• Cell preparation for immunocytochemistry:

  • Grow cells on coated coverslips or chamber slides

  • Fix with 4% paraformaldehyde (10-15 minutes at room temperature)

  • Permeabilize with 0.1-0.5% Triton X-100 in PBS (5-10 minutes)

  • Store in PBS at 4°C if not proceeding immediately

  • Process all experimental conditions simultaneously to ensure comparability

• Sample denaturation for SDS-PAGE:

  • Mix samples with Laemmli buffer containing 5% β-mercaptoethanol

  • Heat to 95°C for 5 minutes to ensure complete denaturation

  • Cool samples to room temperature before loading

  • Include molecular weight markers appropriate for 66-71 kDa range

  • Load 20-40 μg total protein per lane for cell/tissue lysates

These optimized sample preparation methods ensure consistent and reproducible detection of C19orf44 across different experimental platforms while minimizing artifacts and false negative results.

How does antibody validation differ between various applications for C19orf44 detection?

Application-specific validation strategies are essential when using C19orf44 antibodies across different experimental platforms:

• Western Blot validation:

  • Confirm single band at expected molecular weight (66-71 kDa)

  • Test gradient of antibody dilutions (1:200-1:2000)

  • Include positive control samples (L02 cells, mouse liver, rat brain)

  • Perform loading control normalization

  • Consider knockout/knockdown controls if available

  • Validate lot-to-lot consistency with reference samples

• Immunohistochemistry validation:

  • Optimize antigen retrieval methods (citrate vs. EDTA buffer)

  • Test range of antibody dilutions (1:200-1:500)

  • Include positive and negative tissue controls in each run

  • Perform peptide competition assays to confirm specificity

  • Compare staining patterns with mRNA expression data

  • Evaluate subcellular localization consistency

• Immunocytochemistry validation:

  • Compare different fixation methods (PFA, methanol, acetone)

  • Optimize permeabilization conditions

  • Include blocking peptide controls

  • Correlate with Western blot expression in same cell lines

  • Confirm subcellular localization with organelle markers

  • Test signal specificity after siRNA knockdown

• ELISA validation:

  • Generate standard curves with recombinant protein

  • Determine assay detection limits and dynamic range

  • Assess intra- and inter-assay coefficients of variation

  • Confirm sample matrix compatibility

  • Evaluate recovery of spiked standards

  • Test linearity of dilution across sample types

This application-specific validation approach ensures that C19orf44 antibodies perform optimally across different experimental contexts, enhancing data reliability and reproducibility.

What strategies can be employed for multiplexed detection of C19orf44 with other proteins?

Implementing multiplexed detection of C19orf44 alongside other proteins requires these methodological considerations:

• Fluorescence multiplexing in microscopy:

  • Select primary antibodies from different host species (rabbit anti-C19orf44 with mouse/goat antibodies against other targets)

  • Use spectrally distinct fluorophores with minimal overlap

  • Implement sequential staining for antibodies from same species

  • Apply appropriate controls for each antibody individually

  • Consider tyramide signal amplification for low abundance targets

  • Utilize spectral unmixing for closely overlapping fluorophores

• Multiplex immunoblotting approaches:

  • Sequential probing with stripping between antibodies

  • Simultaneous detection using differentially labeled secondary antibodies

  • Use of size-separated proteins (C19orf44 at 66-71 kDa )

  • Digital multiplex Western blot platforms

  • Fluorescence-based Western detection systems

  • Consider protein molecular weight when selecting combinations

• Mass cytometry/Imaging mass cytometry:

  • Metal-conjugated antibodies for highly multiplexed detection

  • Validate each antibody individually before multiplexing

  • Implement comprehensive compensation controls

  • Ensure antibody stability after metal conjugation

  • Apply appropriate clustering and visualization algorithms

  • Use dimensionality reduction techniques for data interpretation

• Proximity ligation assays:

  • Combine C19orf44 antibody with antibodies against interaction partners

  • Optimize primary antibody concentrations individually

  • Include appropriate negative controls (non-interacting protein pairs)

  • Consider dual recognition PLA to enhance specificity

  • Implement quantitative analysis of interaction signals

• Multiplexed immunoassays (Luminex/MSD):

  • Develop capture and detection antibody pairs

  • Validate assay specificity with recombinant proteins

  • Determine cross-reactivity profiles

  • Optimize sample dilution to ensure detection within linear range

  • Include standard curves for each target protein

These methodological approaches enable simultaneous analysis of C19orf44 alongside other proteins of interest, facilitating studies of coexpression patterns, protein interactions, and pathway analyses within the same samples.

How should researchers approach epitope mapping for C19orf44 antibodies?

For comprehensive epitope mapping of C19orf44 antibodies, researchers should implement this systematic approach:

• In silico analysis:

  • Analyze the immunogen sequence used for antibody generation (e.g., Accession Number: BC027869)

  • Perform computational prediction of potential antigenic regions

  • Assess conservation across species for cross-reactivity prediction

  • Evaluate protein secondary structure and accessibility

  • Identify potential post-translational modification sites that might affect binding

• Peptide array mapping:

  • Generate overlapping peptide series spanning C19orf44 sequence

  • Synthesize peptides of 15-20 amino acids with 5-10 amino acid overlap

  • Immobilize peptides on membrane or microarray format

  • Probe with C19orf44 antibody followed by appropriate detection

  • Identify positive signals corresponding to specific peptide sequences

• Deletion/mutation mapping:

  • Generate truncated or domain-specific recombinant C19orf44

  • Express point mutations at predicted antibody binding sites

  • Test antibody reactivity against mutant proteins by Western blot

  • Compare binding affinities using ELISA or surface plasmon resonance

  • Correlate structural changes with binding efficiency

• Cross-reactivity profiling:

  • Test reactivity against related proteins or homologs

  • Evaluate species cross-reactivity experimentally

  • Perform competition assays with related peptides/proteins

  • Determine minimal epitope length required for binding

  • Assess impact of adjacent sequences on binding affinity

• Conformational epitope analysis:

  • Compare native vs. denatured protein recognition

  • Utilize hydrogen-deuterium exchange mass spectrometry

  • Apply chemical crosslinking followed by mass spectrometry

  • Consider crystallography of antibody-antigen complexes

  • Use circular dichroism to assess structural changes upon binding

This systematic epitope mapping approach provides critical information about antibody specificity, potential cross-reactivity, and optimal application conditions, ultimately enhancing the reliability of experimental results with C19orf44 antibodies.

What considerations should be made when selecting between different C19orf44 antibody clones for specific research applications?

When selecting between different C19orf44 antibody clones for specific research applications, consider these critical factors:

• Epitope specificity and location:

  • Determine the antibody's target epitope region when available

  • Consider epitope conservation across species for comparative studies

  • Evaluate whether epitopes are sensitive to fixation/denaturation

  • Assess potential masking by protein-protein interactions

  • Select epitopes appropriate for the biological question (functional domains vs. general detection)

• Validation status for specific applications:

  • Review manufacturer validation data for your intended application

  • Some antibodies are specifically validated for Western blot (1:500-1:1000)

  • Others have validated IHC applications (1:200-1:500)

  • Consider antibodies with multi-application validation when possible

  • Evaluate the extent of validation (single vs. multiple cell/tissue types)

• Technical specifications and format:

  • Compare host species (most C19orf44 antibodies are rabbit polyclonal)

  • Assess antibody concentration and formulation (typically in PBS with 0.02% sodium azide and 50% glycerol)

  • Consider conjugated versions for direct detection if available

  • Evaluate lot-to-lot consistency reports

  • Review storage requirements and shelf-life data

• Experimental evidence and literature support:

  • Search for published studies using specific antibody catalog numbers

  • Contact authors who have successfully used the antibody

  • Consider publication record as evidence of reliability

  • Evaluate figure quality in published applications

  • Assess whether contradictory results exist with specific clones

• Experimental design compatibility:

  • Consider compatibility with other antibodies for co-staining experiments

  • Evaluate antibody performance in your specific sample types

  • Assess background levels in relevant tissues/cells

  • Consider detection method compatibility

  • Evaluate cost-effectiveness for large-scale studies