C11orf68 Antibody

What is C11orf68 and why is it studied in research?

C11orf68, also known as UPF0696 protein C11orf68, Basophilic Leukemia-Expressed Protein Bles03, or Protein P5326, is a protein encoded by the chromosome 11 open reading frame 68 gene . This protein has garnered research interest due to its potential roles in biological processes yet to be fully characterized. Studies utilizing C11orf68 antibodies help elucidate its expression patterns, subcellular localization, and possible functions in normal and pathological conditions . Methodologically, researchers typically employ these antibodies in protein detection assays such as Western blotting, immunohistochemistry, and immunofluorescence to map expression across tissues and cellular compartments.

What are the different types of C11orf68 antibodies available for research?

C11orf68 antibodies come in several forms differentiated by host organisms, clonality, binding specificity, and conjugation status:

Selection should be based on experimental requirements, with unconjugated antibodies offering flexibility for secondary detection systems, while conjugated versions provide direct visualization or detection capabilities .

What is the species reactivity profile of available C11orf68 antibodies?

Most commercially available C11orf68 antibodies demonstrate reactivity with human samples, which is the primary research focus . Some antibodies offer cross-reactivity with mouse and rat samples, allowing for comparative studies across species models . The antibody cataloged as STJ194102 specifically shows reactivity with human, mouse, and rat samples , while other products like the Proteintech antibody (84725-5-RR) have been validated extensively in human samples including A431 cells, PC-3 cells, and human stomach cancer tissue . When designing cross-species experiments, researchers should carefully verify the specific reactivity claims with validation data from manufacturers.

What are the optimal dilution ratios for different C11orf68 antibody applications?

Dilution optimization is critical for balancing signal strength against background. Based on validated protocols:

Each new antibody lot should undergo titration validation, as optimal concentrations may vary between manufacturers and applications .

How should C11orf68 antibodies be stored and handled to maintain activity?

Proper storage and handling significantly impact antibody performance. For C11orf68 antibodies:

• Store at -20°C or -80°C for long-term preservation

• Avoid repeated freeze/thaw cycles which can cause protein denaturation and reduced activity

• For light-sensitive conjugates like FITC, minimize exposure to light during storage and handling

• Most C11orf68 antibodies are supplied in buffer containing 50% glycerol, allowing for storage at -20°C without freezing solid

• Working aliquots can be stored at 4°C for up to one month, but prolonged storage at this temperature is not recommended

When working with these antibodies, thaw completely before use, mix gently to avoid foaming, and centrifuge briefly to collect all liquid at the bottom of the tube before opening .

What sample preparation methods improve C11orf68 detection in Western blot applications?

Effective sample preparation enhances detection sensitivity and specificity:

• Cell lysis buffers containing protease inhibitors are essential to prevent protein degradation

• For membrane-associated proteins, include detergents like NP-40, Triton X-100, or CHAPS

• Sample denaturation conditions should be optimized (temperature, reducing agents)

• Loading 20-40 μg of total protein typically yields detectable C11orf68 signals

• Fresh samples generally produce cleaner results than frozen-thawed lysates

Based on published protocols, C11orf68 antibodies have successfully detected endogenous levels in human cell lines including A431 and PC-3 . Methodologically, researchers should include positive control samples of known C11orf68 expression to validate detection and quantification procedures.

How can nonspecific binding be minimized when using C11orf68 antibodies?

Nonspecific binding is a common challenge that can obscure meaningful results:

• Optimize blocking conditions using 3-5% BSA or milk in TBST/PBST, testing which gives cleaner background

• For tissue samples, pre-absorb antibodies with tissue powder from a species different from your experimental samples

• Increase washing duration and volume (minimum three 10-minute washes with gentle agitation)

• Include 0.1-0.3% Tween-20 in wash buffers to reduce hydrophobic nonspecific interactions

• For IHC applications with C11orf68 antibodies, consider antigen retrieval with TE buffer at pH 9.0 as recommended for optimal results

Experimentally, comparing binding patterns between related antibodies targeting different epitopes of C11orf68 can help distinguish specific from nonspecific signals.

What controls should be included when validating C11orf68 antibody specificity?

Rigorous validation requires appropriate controls:

• Positive tissue/cell controls: Use samples with known C11orf68 expression (A431 cells, PC-3 cells have been validated)

• Negative controls: Include samples where the target is known to be absent or knocked down

• Peptide competition assays: Pre-incubate antibody with immunizing peptide to confirm specificity

• Secondary-only controls: Omit primary antibody to assess background from secondary detection

• Isotype controls: Use non-specific IgG from the same host species and at the same concentration

• Knockdown/knockout validation: Compare staining in wild-type versus C11orf68-depleted samples

These controls should be performed for each new application or sample type to ensure reliable data interpretation and reproducibility.

How can researchers address discrepancies in C11orf68 detection between different antibodies?

When different antibodies yield conflicting results:

• Compare epitope recognition sites—antibodies targeting different domains may detect different isoforms or post-translationally modified variants

• Evaluate antibody validation data from manufacturers for each specific application

• Consider protein conformation effects—some epitopes may be masked in particular cellular compartments or under certain fixation conditions

• Use orthogonal techniques (e.g., mass spectrometry) to confirm protein identity

• Test multiple antibodies in parallel on the same samples to determine the most consistent performer

For C11orf68 specifically, antibodies targeting the region AA 1-251 versus those targeting AA 151-201 might yield different results depending on protein processing or interaction states .

How should experiments be designed to study C11orf68 expression changes in disease models?

Effective experimental design follows these principles:

• Include sufficient biological replicates (minimum triplicates) to account for natural variation

• Randomize sample preparation to avoid batch effects

• Process control and experimental samples simultaneously to minimize technical variation

• Include technical replicates for validation of detection method consistency

• For differential expression studies, carefully control for confounding variables and consider power analysis for sample size determination

Proteintech's C11orf68 antibody has been validated in human stomach cancer tissue , suggesting potential applications in cancer research where proper experimental controls must include matched normal tissue from the same patients whenever possible.

What are appropriate normalization methods when quantifying C11orf68 protein levels?

Proper normalization is essential for accurate comparative analysis:

• For Western blots:

  • Normalize to housekeeping proteins (β-actin, GAPDH, α-tubulin) after confirming their stability across experimental conditions

  • Consider total protein normalization using stain-free gels or membrane staining as an alternative approach

  • Apply lane normalization to account for loading differences

• For immunohistochemistry:

  • Use digital image analysis with appropriate background subtraction

  • Apply tissue-specific normalization to account for variability in sample composition

  • Consider cell-type specific quantification for heterogeneous tissues

• For all quantitative analyses:

  • Report normalization methods transparently

  • Validate normalization controls across experimental conditions

  • Apply appropriate statistical tests based on data distribution and experimental design

How can C11orf68 antibodies be incorporated into multi-parameter analysis techniques?

Advanced research often requires simultaneous analysis of multiple targets:

• Multiplex immunofluorescence:

  • Combine C11orf68 FITC-conjugated antibodies with other fluorophore-conjugated antibodies against different targets

  • Carefully select fluorophores with minimal spectral overlap

  • Include appropriate controls for each antibody individually

• Flow cytometry applications:

  • C11orf68 FITC-conjugated antibodies can be incorporated into multicolor panels

  • Titrate antibodies in the context of the full panel to account for fluorophore interactions

  • Include fluorescence-minus-one (FMO) controls for accurate gating

• Protein interaction studies:

  • Use C11orf68 antibodies for co-immunoprecipitation followed by mass spectrometry

  • Consider proximity ligation assays to identify interaction partners in situ

  • Validate interactions with reciprocal immunoprecipitation experiments

These multiparameter approaches require careful optimization of antibody concentrations and detection parameters to ensure reliable and reproducible results.

How can C11orf68 antibodies contribute to understanding protein function in translational research?

C11orf68 represents a protein with potential undiscovered functions that antibody-based research can help elucidate:

• Tissue expression mapping:

  • Systematic analysis across normal and pathological human tissues

  • Correlation with clinical outcomes in disease states

  • Development of diagnostic or prognostic biomarkers

• Subcellular localization studies:

  • High-resolution imaging with validated C11orf68 antibodies

  • Changes in localization during cellular processes or stress conditions

  • Co-localization with known functional protein complexes

• Functional analyses:

  • Combine antibody-based detection with knockout/knockdown approaches

  • Assess effects on cellular pathways through phospho-protein analysis

  • Identify differential expression in response to therapeutic interventions

The validated reactivity of current antibodies with human, mouse, and rat samples enables translational research spanning from basic mechanisms to potential clinical applications.

What technical considerations are important when integrating C11orf68 antibody data with genomic expression analyses?

Multi-omics integration requires careful technical approaches:

• When correlating protein and mRNA levels:

  • Account for potential post-transcriptional regulation

  • Consider temporal differences between transcription and translation

  • Use statistical methods appropriate for integrating different data types

• For single-cell applications:

  • Validate antibody specificity in low-abundance situations

  • Develop protocols that preserve both RNA and protein epitopes

  • Apply computational approaches that can handle multi-modal data

• Technical validation:

  • Confirm antibody detection correlates with transcript abundance in controlled systems

  • Account for protein stability and half-life when comparing to mRNA data

  • Consider alternative splicing events that might affect epitope presence

Understanding these technical limitations is essential for meaningful integration of antibody-based protein detection with transcriptomic data, particularly when studying proteins like C11orf68 where functional characterization is still emerging.