KCNMB3 Antibody, HRP conjugated

What specific applications has KCNMB3 Antibody, HRP conjugated been validated for?

The KCNMB3 Antibody, HRP conjugated (CSB-PA878838LB01HU) has been specifically tested and validated for ELISA applications . Other non-HRP conjugated KCNMB3 antibodies have broader application profiles including Western Blotting (WB), Immunohistochemistry (IHC), and Flow Cytometry . When designing experiments, it's important to note that validated applications vary between conjugated and unconjugated versions of the antibody.

For optimal results in ELISA applications:

• Use freshly prepared samples and reagents

• Follow recommended dilution guidelines (specific to each antibody lot)

• Include appropriate positive and negative controls

• Consider using blocking buffers containing 1-5% BSA to minimize background

What are the recommended storage conditions for maintaining KCNMB3 Antibody, HRP conjugated activity?

According to manufacturer specifications, KCNMB3 Antibody, HRP conjugated should be stored at -20°C or -80°C upon receipt . Researchers should:

• Avoid repeated freeze-thaw cycles which can compromise antibody activity and epitope recognition

• Store in small aliquots (typically 10-50 μL) to minimize freeze-thaw events

• Keep the antibody in its storage buffer (50% Glycerol, 0.01M PBS, pH 7.4 with 0.03% Proclin 300 as preservative)

• Monitor storage temperature consistency, as temperature fluctuations can reduce shelf life

• For short-term use (within 1 week), storage at 4°C is acceptable

How should researchers design validation experiments for KCNMB3 Antibody, HRP conjugated?

Validation experiments should include:

• Positive and negative controls:

  • Positive: Human brain tissue or cell lines known to express KCNMB3

  • Negative: Tissues with minimal KCNMB3 expression or KCNMB3 knockout samples

• Cross-reactivity assessment:

  • Test against related potassium channel proteins (KCNMB1, KCNMB2, KCNMB4)

  • Evaluate species cross-reactivity (antibody was raised against human KCNMB3)

• Peptide competition assays:

  • Pre-incubate antibody with immunizing peptide (residues 82-207 of human KCNMB3)

  • Compare signal with and without peptide competition

• Dilution optimization:

  • Create a dilution series to determine optimal working concentration

  • Start with manufacturer's recommended range and adjust based on signal-to-noise ratio

How can researchers distinguish between different KCNMB3 isoforms?

KCNMB3 undergoes alternative splicing, resulting in multiple isoforms with distinct functional properties . To distinguish between isoforms:

• Epitope mapping:

  • Determine which protein region (residues 82-207) the antibody recognizes

  • Compare this with known splicing regions in KCNMB3

• Molecular weight analysis:

  • KCNMB3 has a calculated molecular weight of approximately 32 kDa

  • Different isoforms exhibit slight variations in electrophoretic mobility

  • Isoform 1, 2, 3, and 4 show distinct functional properties in electrophysiological studies

• Functional correlation:

  • Isoform 2, 3, and 4 partially inactivate KCNMA1 currents

  • Isoform 1 does not induce detectable inactivation of KCNMA1

  • Isoform 4 induces fast but incomplete inactivation detectable only at large depolarizations

What are the most common technical challenges when using KCNMB3 Antibody, HRP conjugated, and how can they be addressed?

Common challenges include:

• High background signal:

  • Increase blocking time (2-3 hours at room temperature)

  • Use 3-5% BSA in TBS-T for blocking

  • Optimize antibody dilution (typically 1:500-1:2000 for related KCNMB3 antibodies)

  • Include 0.05% Tween-20 in wash buffers

• Weak or no signal:

  • Verify KCNMB3 expression in sample (membrane protein extraction may require specialized protocols)

  • Increase antibody concentration or incubation time

  • Use enhanced detection systems such as high-sensitivity ECL substrates

  • Confirm HRP activity has not been compromised during storage

• Non-specific bands:

  • Increase stringency of washing steps (more washes, longer duration)

  • Pre-adsorb antibody with non-specific proteins

  • Use gradient gels to improve separation of similar molecular weight proteins

What methodological modifications are necessary when working with tissue samples versus cell cultures?

How can KCNMB3 Antibody, HRP conjugated be utilized in studying the relationship between potassium channels and disease models?

Researchers can employ this antibody to investigate:

• Hypertension models:

  • While KCNMB3 itself hasn't been directly linked to hypertension, related potassium channel genes (KCNK3, KCNK9) have demonstrated associations with blood pressure regulation and aldosterone production

  • Investigate KCNMB3 expression in models of hyperaldosteronism

  • Correlate KCNMB3 activity with electrophysiological parameters in cardiovascular tissues

• Neurological disorders:

  • MaxiK channels are fundamental to neuronal excitability

  • Study KCNMB3 expression in neurological disease models

  • Investigate interactions between KCNMB3 and alpha subunits in neural tissues

  • Explore the role of different isoforms in modulating neuronal firing patterns

• Cancer research applications:

  • Following the model of KCNAB2 in lung cancer research , investigate KCNMB3 expression in various cancer types

  • Analyze correlations between KCNMB3 expression and tumor progression

  • Explore potential diagnostic or prognostic value

What are the considerations when integrating KCNMB3 Antibody, HRP conjugated into multi-protein complex studies?

When investigating KCNMB3 in the context of multi-protein complexes:

• Co-immunoprecipitation optimizations:

  • Select lysis buffers that preserve membrane protein interactions

  • Consider mild detergents (0.5-1% NP-40 or digitonin) rather than harsh detergents (SDS)

  • Adjust salt concentration to maintain complex integrity

  • Include phosphatase/protease inhibitors to preserve post-translational modifications

• BK channel tetramer studies:

  • Two or more KCNMB3 subunits are required to block the KCNMA1 tetramer

  • Design experiments to assess stoichiometry of alpha-beta interactions

  • Consider native PAGE or crosslinking approaches to preserve complex structure

• Functional correlations:

  • Combine antibody detection with electrophysiological measurements

  • Correlate KCNMB3 detection with calcium sensitivity measurements

  • Integrate findings with patch-clamp data to correlate protein expression with channel function

How does KCNMB3 Antibody, HRP conjugated compare with other molecular tools for studying potassium channels?

What methodological approaches should be considered when studying KCNMB3 interactions with the KCNMA1 alpha subunit?

To investigate the regulatory interactions between KCNMB3 and KCNMA1:

• Proximity ligation assays:

  • Detect direct protein-protein interactions in situ

  • Require separate antibodies against KCNMB3 and KCNMA1

  • Provide spatial information about interaction sites

• Co-expression studies:

  • Express tagged versions of KCNMB3 and KCNMA1

  • Use the HRP-conjugated antibody to detect KCNMB3 in co-expression systems

  • Correlate with electrophysiological changes in channel activity

  • Assess how different KCNMB3 isoforms modulate KCNMA1 calcium sensitivity and gating kinetics

• Biochemical characterization:

  • Study how KCNMB3 alters the biophysical properties of KCNMA1

  • Investigate the stoichiometry of alpha-beta subunit interaction

  • Determine how two or more KCNMB3 subunits block the KCNMA1 tetramer

  • Use crosslinking approaches to stabilize the channel complex