ppk15 Antibody

Basic Research Questions

• What is PPK15 antibody and what is its target specificity?

  PPK15 antibody is a polyclonal antibody raised against Schizosaccharomyces pombe (fission yeast) PPK15 protein. According to product specifications, it is a rabbit polyclonal antibody generated using recombinant S. pombe PPK15 protein as the immunogen . This antibody recognizes the native PPK15 protein, which functions in cellular signaling pathways in fission yeast systems. When selecting this antibody for experiments, researchers should carefully consider the specific epitope recognition and validate cross-reactivity if working with related proteins or orthologous targets.

• What applications has PPK15 antibody been validated for?

  The PPK15 antibody has been primarily validated for ELISA and Western blot applications . For Western blotting, the recommended concentration range is 1.0-10 μg/ml, though optimization for specific experimental conditions is advisable. When using this antibody in these applications, researchers should include appropriate controls to ensure specificity and reliability of results. Although not explicitly validated for other techniques, researchers may test its utility in additional applications following proper validation protocols as outlined in antibody validation literature .

• How should PPK15 antibody be validated before experimental use?

  Proper validation should follow a multi-step approach:

  • Specificity testing: Perform Western blot analysis to confirm single band detection at the expected molecular weight (~74 kDa for typical target proteins)

  • Positive and negative controls: Include samples known to express or lack the target protein

  • Blocking peptide experiments: If available, use competing peptides to confirm epitope specificity

  • Knockout/knockdown validation: Test in systems where the target has been genetically removed or reduced

  • Cross-application validation: Compare results across multiple techniques (e.g., WB, ELISA, IF)

  According to antibody validation literature, these steps help ensure that the observed signal is specific to the target protein and not due to non-specific binding or cross-reactivity .

• What are the recommended storage and handling conditions for PPK15 antibody?

  Based on manufacturer specifications for similar antibodies, optimal storage conditions include:

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

  • Avoid repeated freeze-thaw cycles to maintain antibody functionality

  • Store in the supplied buffer containing 50% glycerol and 0.01M PBS (pH 7.4) with preservative

  • When working with the antibody, keep on ice and return to storage promptly

  • For long-term stability, consider preparing working aliquots to minimize freeze-thaw cycles

  Proper storage significantly affects antibody performance and reproducibility across experiments, particularly for polyclonal antibodies that may contain diverse epitope recognition .

• What controls should be included when using PPK15 antibody in experiments?

  Essential controls include:

  • Positive control: Sample known to express the target protein

  • Negative control: Sample lacking the target protein (ideally knockout or naturally negative)

  • Technical control: Omitting primary antibody to assess secondary antibody specificity

  • Loading control: For Western blots, include housekeeping protein detection

  • Blocking peptide control: When available, pre-incubate antibody with immunizing peptide

  These controls help distinguish between specific and non-specific signals and validate experimental results, particularly important for polyclonal antibodies which may have batch-to-batch variation .