KCNK15 Antibody,HRP conjugated

What is KCNK15 and what biological functions does it serve?

KCNK15 (Potassium channel subfamily K member 15) is a probable potassium channel subunit, also known as TASK-5 (TWIK-related acid-sensitive K(+) channel 5). It belongs to the two-pore potassium channel family. Although no channel activity has been observed in heterologous systems, research suggests it may need to associate with another protein to form a functional channel . KCNK15 is involved in potassium ion transport across cellular membranes and may play roles in membrane excitability and cellular homeostasis.

The protein functions in the context of signal transduction pathways , making it relevant to neurological, cardiac, and oncological research. Current evidence indicates KCNK15 has potential significance in cancer biology, particularly in thyroid and lung cancers .

What are the specifications and optimal storage conditions for KCNK15 Antibody,HRP conjugated?

KCNK15 Antibody,HRP conjugated is typically a rabbit polyclonal antibody targeting human KCNK15. The key specifications include:

For optimal preservation of antibody activity:

• Store at -20°C or -80°C upon receipt

• Avoid repeated freeze-thaw cycles

• Consider aliquoting the antibody for long-term storage to minimize freeze-thaw cycles

What applications is KCNK15 Antibody,HRP conjugated suitable for?

When transitioning between applications, researchers should validate the antibody's performance in their specific experimental system .

What controls should be included when using KCNK15 Antibody,HRP conjugated in research?

For rigorous experimental design with KCNK15 Antibody,HRP conjugated:

• Positive Control: Include a sample known to express KCNK15 (e.g., relevant human cancer cell lines based on research context)

• Negative Control:

  • Primary antibody omission

  • Use of isotype control (rabbit IgG is the appropriate control)

  • Samples from KCNK15 knockdown/knockout models

• Blocking Peptide Control: When available, using the immunizing peptide to pre-absorb the antibody can confirm specificity

• Cross-Reactivity Considerations: If working with non-human samples, note the predicted reactivity percentages: Cow: 83%, Dog: 93%, Human: 100%, Pig: 92%

• Secondary Antibody Controls: For non-conjugated versions requiring secondary antibodies, include secondary-only controls

How can I optimize ELISA protocols using KCNK15 Antibody,HRP conjugated?

For optimal ELISA performance with KCNK15 Antibody,HRP conjugated:

• Dilution Optimization: Start with the recommended 1:20000 dilution , but perform a titration experiment (1:10000 to 1:40000) to determine optimal signal-to-noise ratio for your specific samples

• Blocking Protocol: Use 5% non-fat dry milk or 3% BSA in PBS-T to minimize non-specific binding

• Sample Preparation:

  • For cell lysates: Use a buffer containing protease inhibitors

  • For tissue samples: Homogenize thoroughly and ensure uniform protein concentration

• Incubation Conditions:

  • Temperature: Perform primary antibody incubation at 4°C overnight for maximum specificity

  • Washing: Include at least 3-5 washes with PBS-T between steps to reduce background

• Signal Development:

  • Since the antibody is HRP-conjugated, use appropriate substrates like TMB

  • Monitor color development to avoid saturation

  • Consider measuring at multiple timepoints for optimal signal

• Troubleshooting:

  • High background: Increase dilution or blocking time

  • Weak signal: Decrease dilution or increase sample concentration

How can KCNK15 Antibody,HRP conjugated be used to study the role of KCNK15 in thyroid cancer?

Research has shown significant upregulation of KCNK15 mRNA expression in thyroid cancer tissues compared to normal tissues . To investigate this association:

• Expression Analysis:

  • Use KCNK15 Antibody in ELISA to quantify expression levels in thyroid cancer patient samples versus normal controls

  • Compare expression across different cancer stages to validate the correlation between KCNK15 expression and tumor stage reported in literature

• Prognostic Value Assessment:

  • Design studies correlating KCNK15 protein levels with patient survival data

  • Implement multivariate analyses to evaluate KCNK15 as an independent prognostic marker

• Mechanistic Studies:

  • Combine with functional assays after KCNK15 knockdown/overexpression

  • Integrate with analysis of related KCNK family members (KCNK2, KCNK4, KCNK5) that have also been implicated in thyroid cancer

• Biomarker Development:

  • Evaluate KCNK15 antibody performance in multiplex assays with other thyroid cancer markers

  • Develop standardized ELISA protocols for potential clinical application

• Therapeutic Target Validation:

  • Use in drug screening assays targeting KCNK15

  • Monitor KCNK15 expression changes in response to various treatment modalities

What is the relationship between KCNK15, KCNK15-AS1, and cancer progression?

While KCNK15 is the protein target of the antibody, research has revealed intriguing connections with KCNK15-AS1 (antisense RNA) in cancer:

• Dual Expression Analysis:

  • KCNK15-AS1 is significantly overexpressed in lung adenocarcinoma tissues

  • High expression of KCNK15-AS1 correlates with poor prognosis, poor differentiation, advanced clinical stage, and lymph node metastasis

• Functional Relationship:

  • KCNK15-AS1 knockdown inhibits lung cancer cell proliferation

  • This effect is partially mediated through miR-202/miR-370/EGFR signaling axis

• Research Application Strategy:

  • Use KCNK15 Antibody,HRP conjugated alongside RNA detection methods for KCNK15-AS1

  • Investigate potential regulatory relationships between the antisense RNA and protein expression

  • Examine correlation between KCNK15 protein levels and KCNK15-AS1 expression in the same samples

• Signaling Pathway Analysis:

  • Explore whether KCNK15 protein function is affected by KCNK15-AS1 expression

  • Determine if KCNK15-AS1 functions independently of KCNK15 protein

What methodological approaches can resolve conflicting data between mRNA and protein expression of KCNK15?

When investigating discrepancies between KCNK15 mRNA and protein expression:

• Multi-level Validation:

  • Perform parallel qRT-PCR and ELISA/Western blot on the same samples

  • Include multiple primer sets and antibodies targeting different epitopes

  • Use tissue microarrays with matched normal/tumor samples for large-scale validation

• Post-transcriptional Regulation Assessment:

  • Investigate miRNA targeting of KCNK15 mRNA

  • Examine KCNK15 mRNA stability using actinomycin D chase experiments

  • Analyze polysome profiles to assess translational efficiency

• Protein Stability Analysis:

  • Perform cycloheximide chase experiments to determine KCNK15 protein half-life

  • Investigate proteasomal and lysosomal degradation pathways

  • Assess post-translational modifications that might affect antibody recognition

• Subcellular Localization Studies:

  • Use cell fractionation followed by Western blot

  • Perform immunocytochemistry to determine if protein is sequestered in specific compartments

  • Consider native versus denatured protein detection methods

• Cross-platform Comparison:

  • Compare RNA-seq data with proteomics results

  • Utilize the TCGA and CPTAC databases for integrated analysis

How can I validate the specificity of KCNK15 Antibody,HRP conjugated?

To establish confidence in antibody specificity:

• Epitope Analysis:

  • Compare the immunogen sequence (amino acids 244-330AA) with other proteins to identify potential cross-reactivity

  • Note that some antibodies target more specific regions like the C-terminal (273-322) or middle region

• Knockout/Knockdown Validation:

  • Generate KCNK15 knockdown using siRNA or CRISPR-Cas9

  • Confirm signal reduction/elimination in knockout samples

• Overexpression Studies:

  • Express tagged KCNK15 in a system with low endogenous expression

  • Confirm co-localization of tag and antibody signals

• Peptide Competition:

  • Pre-incubate antibody with excess immunizing peptide

  • Verify signal abolishment in subsequent assays

• Cross-Reactivity Assessment:

  • Test on samples from multiple species if relevant

  • Consider predicted reactivity percentages: Cow (83%), Dog (93%), Human (100%), Pig (92%)

What factors might affect KCNK15 detection in experimental samples?

Several biological and technical factors can impact detection:

• Sample Preparation Issues:

  • Insufficient protein extraction from membrane fractions (KCNK15 is a membrane protein)

  • Protein degradation during isolation

  • Epitope masking due to fixation conditions (especially for IHC)

• Experimental Variables:

  • Buffer composition affecting antibody binding

  • Presence of detergents that might denature the epitope

  • pH conditions altering antibody-antigen interaction

• Biological Variability:

  • Post-translational modifications affecting epitope recognition

  • Alternative splicing generating isoforms not recognized by the antibody

  • Protein-protein interactions masking antibody binding sites

• Technical Recommendations:

  • Use multiple extraction methods to ensure comprehensive protein isolation

  • Include protease and phosphatase inhibitors in all buffers

  • Consider native versus denaturing conditions depending on epitope location

How might KCNK15 Antibody,HRP conjugated contribute to understanding potassium channel dysregulation in disease?

Emerging research opportunities include:

• Cancer Biology Applications:

  • Explore KCNK15 as part of the potassium channel dysregulation signature in multiple cancer types beyond thyroid cancer

  • Investigate correlation between KCNK15 expression and response to ion channel-targeting drugs

  • Develop combination biomarker panels including multiple KCNK family members

• Signaling Pathway Integration:

  • Map the relationship between KCNK15 expression and major oncogenic pathways

  • Investigate KCNK15 regulation by growth factors and cellular stress

  • Explore potential roles in cell migration and invasion processes

• Therapeutic Target Validation:

  • Screen compounds specifically targeting KCNK15

  • Evaluate KCNK15 as a predictive marker for response to existing therapies

  • Investigate combinatorial approaches targeting multiple ion channels

• Clinical Translation Potential:

  • Develop standardized ELISA protocols for potential diagnostic applications

  • Explore KCNK15 in liquid biopsy approaches

  • Correlate KCNK15 levels with treatment response and disease progression

What methodological innovations might enhance research using KCNK15 antibodies?

Future technological developments could include:

• Advanced Conjugation Options:

  • Expanding beyond HRP to fluorescent conjugates for multicolor flow cytometry

  • Developing antibody fragments for improved tissue penetration

  • Creating bispecific antibodies targeting KCNK15 and relevant signaling partners

• Single-Cell Applications:

  • Adapting KCNK15 antibodies for mass cytometry (CyTOF)

  • Developing protocols for single-cell Western blotting

  • Integrating with spatial transcriptomics technologies

• Functional Assays:

  • Developing antibodies that modulate channel function

  • Creating toolkits for measuring KCNK15 channel activity in live cells

  • Combining with electrophysiology for structure-function studies

• Clinical Research Applications:

  • Standardizing KCNK15 detection for potential companion diagnostic development

  • Creating multiplex assays including multiple KCNK family members

  • Developing automation-compatible high-throughput protocols