LSM7 Antibody

Here’s a structured FAQ collection for LSM7 antibody research, organized by methodological complexity and supported by experimental evidence from peer-reviewed studies:

How do I validate the specificity of an LSM7 antibody in Western blot (WB) assays?

• Methodology:

  • Use positive controls (e.g., HeLa cell lysates) to confirm antibody binding at ~12 kDa, matching LSM7’s predicted molecular weight .

  • Include knockout controls (e.g., CRISPR/Cas9-mediated LSM7 KO cell lines) to verify band absence .

  • Compare results across multiple antibodies (e.g., Proteintech 18941-1-AP vs. Aviva ARP40837_P050) to rule out non-specific binding .

What are optimal dilution protocols for LSM7 antibodies in immunohistochemistry (IHC)?

• Guidelines:

  • Start with 1:20–1:200 dilution for IHC in human tissues, using TE buffer (pH 9.0) for antigen retrieval .

  • Titrate in pilot experiments to balance signal-to-noise ratios, especially in dense tissues (e.g., brain or lymphoma samples) .

How do LSM7 antibody performance issues arise in phase-separation studies, and how can they be resolved?

• Problem: LSM7 forms stress granule (SG) condensates via liquid-liquid phase separation (LLPS), but antibodies may fail to detect dynamic aggregates .

• Solutions:

  • Use fresh lysates with protease/phosphatase inhibitors to preserve post-translational modifications.

  • Combine immunofluorescence with LSM7-GFP tagging to confirm localization .

  • Validate with LSM7 deletion strains (e.g., yeast lsm7Δ) to distinguish background noise .

How to reconcile contradictory data on LSM7’s role in oligodendrocyte survival versus stress granule formation?

• Context:

  • Study 1: LSM7 knockdown in zebrafish increased oligodendrocyte apoptosis .

  • Study 2: Yeast Lsm7 promotes SG assembly via phase separation .

• Resolution framework:

  • Assess tissue-specific isoforms (human vs. yeast LSM7 share 83% homology) .

  • Use cell-type-specific models (e.g., oligodendrocyte cultures vs. yeast).

  • Monitor RNA-binding dynamics: LSM7’s U6 snRNA association may differ across species .

What strategies ensure reproducibility of LSM7 antibody results across laboratories?

• Best practices:

  • Standardize lysate preparation (e.g., RIPA buffer with 1% SDS for WB) .

  • Share positive/negative controls (e.g., human lymphoma tissue for IHC) .

  • Use publicly available datasets (e.g., PMC8756503’s ΔΔCt normalization methods) .

How to address cross-reactivity concerns when using LSM7 antibodies in non-model organisms?

• Approach:

  • Validate homology: Zebrafish Lsm7 shares 86% sequence identity with human LSM7 .

  • Pre-adsorb antibodies with target peptides (e.g., Aviva’s AAP40837 blocking peptide) .

  • Perform parallel experiments in established models (e.g., mouse vs. zebrafish) .