SPCC622.14 Antibody

What is the molecular target of SPCC622.14 Antibody and its cellular distribution?

SPCC622.14 Antibody targets cytokeratin 14, a type I intermediate filament protein predominantly expressed in stratifying epithelial cells. This specific marker is crucial for distinguishing stratifying epithelial cells from simple epithelial cells, as the latter do not express cytokeratin 14 . The antibody recognizes the last 15 C-terminal residues of human cytokeratin 14, typically conjugated to thyroglobulin as the immunogen . At the subcellular level, cytokeratin 14 is associated with multiple cellular compartments including the cytosol, nucleus, and mitochondria, while functionally participating in keratin filament formation and hemidesmosome assembly .

What species cross-reactivity has been validated for SPCC622.14 Antibody?

It is important to note that antibody reactivity and optimal working conditions may vary substantially between species, requiring protocol optimization for each specific application and organism .

What validated applications have been established for SPCC622.14 Antibody?

SPCC622.14 Antibody has been validated for multiple experimental applications. The following table outlines verified applications with recommended working dilutions:

For flow cytometry applications, membrane permeabilization is required, with Leucoperm recommended as the permeabilization agent. For paraffin-embedded sections, antigen retrieval using heat treatment with sodium citrate buffer (pH 6.0) is necessary for optimal results .

How should storage and handling protocols be modified to maintain SPCC622.14 Antibody activity?

SPCC622.14 Antibody requires specific storage and handling protocols to maintain its activity and specificity:

• Upon receipt, aliquot the antibody to minimize freeze-thaw cycles

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

• Keep working aliquots at 2-8°C for short-term use (up to 4 weeks)

• Avoid repeated freezing and thawing as this may denature the antibody

• Storage in frost-free freezers is not recommended due to temperature fluctuations

The purified IgG liquid formulation typically contains phosphate-buffered saline with 0.09% sodium azide as a preservative. When properly stored and handled, the antibody maintains guaranteed activity for 12 months from the date of dispatch .

How can SPCC622.14 Antibody be implemented in multi-parameter flow cytometry studies?

For multi-parameter flow cytometry applications, SPCC622.14 Antibody requires specific optimization steps:

• Membrane permeabilization protocol: Since cytokeratin 14 is an intracellular protein, complete membrane permeabilization is essential. Use 10μl of the suggested working dilution (1:100) to label 1×10^6 cells in 100μl of permeabilization buffer .

• Co-staining considerations: When designing multi-parameter panels, consider fluorophore selection to minimize spectral overlap with other markers. Reference publications by Clark et al. (2011) have successfully implemented multi-parameter approaches for molecular subtyping .

• Validation controls: Include both positive controls (stratified epithelial tissues such as skin) and negative controls (simple epithelial tissues) in each experiment to confirm antibody specificity and performance .

• Data analysis strategy: Implement hierarchical gating strategies that first identify epithelial populations before assessing cytokeratin 14 expression to improve signal discrimination.

What methodological approaches should be considered when using SPCC622.14 Antibody for tumor characterization studies?

When implementing SPCC622.14 Antibody for tumor characterization, several methodological considerations are critical:

• Tissue preparation: For optimal results in paraffin-embedded tumor samples, heat-induced antigen retrieval using sodium citrate buffer (pH 6.0) is essential to expose epitopes that may be masked during fixation .

• Comparative analysis: Cytokeratin 14 expression patterns should be analyzed in conjunction with other basal cell markers. Studies by Wetzels et al. (1989) demonstrated the value of combining basement membrane components detection with cytokeratin 14 analysis to distinguish between noninvasive and invasive breast carcinomas .

• Quantitative assessment: Implement digital image analysis to quantify staining intensity and distribution patterns, particularly when evaluating heterogeneous expression in tumor samples as described in molecular subtyping studies by Clark et al. (2011) .

• Research methodology framework: Adopt a mixed-method research approach that combines quantitative assessment of cytokeratin 14 expression with qualitative morphological evaluation for comprehensive tumor characterization .

What are the common technical challenges when using SPCC622.14 Antibody and their solutions?

How can researchers validate the specificity of SPCC622.14 Antibody in novel experimental systems?

Validating SPCC622.14 Antibody specificity in novel experimental systems requires a systematic approach:

• Positive and negative control tissues: Include skin as a positive control (known to express cytokeratin 14) and simple epithelial tissues as negative controls in each experiment .

• Western blot validation: Perform Western blot analysis to confirm the antibody recognizes a protein of the expected molecular weight (approximately 50 kDa for cytokeratin 14). Previous studies (Alam et al., 2011) have validated this application .

• Antibody titration: Conduct a dilution series experiment to determine the optimal antibody concentration that maximizes specific signal while minimizing background .

• Knockout/knockdown controls: When available, include samples from cytokeratin 14 knockout or knockdown systems to confirm signal specificity.

• Research methodology documentation: Thoroughly document all validation steps according to scientific research methodology standards to ensure reproducibility and reliability of findings .

How should SPCC622.14 Antibody staining patterns be interpreted in the context of tissue heterogeneity?

Interpreting SPCC622.14 Antibody staining patterns requires careful consideration of tissue context and cellular heterogeneity:

What emerging applications are being developed for SPCC622.14 Antibody in stem cell research?

SPCC622.14 Antibody has demonstrated valuable applications in stem cell research, particularly in identifying and characterizing epithelial stem cell populations:

• Prostatic epithelial stem cell identification: Richardson et al. (2004) utilized cytokeratin 14 antibodies in conjunction with CD133 to identify and characterize prostatic epithelial stem cells .

• Tumor-initiating cell characterization: Collins et al. (2005) employed cytokeratin 14 detection to prospectively identify tumorigenic prostate cancer stem cells, demonstrating the antibody's utility in cancer stem cell research .

• Regenerative medicine applications: Takahashi et al. (2010) used cytokeratin 14 antibodies to characterize newly established cell lines from mouse oral epithelium capable of regenerating teeth when combined with dental mesenchyme .

• Methodological integration: These applications require integrating cytokeratin 14 detection into comprehensive research methodologies that include multiple stem cell markers and functional assays .