LCMT1 Antibody, FITC conjugated

Key Properties

Recommended Usage

Data inferred from unconjugated LCMT1 antibody protocols ; FITC-conjugated variants typically follow similar optimization.

Biological Relevance of LCMT1

LCMT1’s enzymatic activity is essential for PP2A/PP4 complex assembly and function:

• PP2A Methylation: LCMT1 methylates the C-terminal leucine of PP2A catalytic subunits (PP2Ac), enabling stable binding of regulatory B subunits (e.g., Bα, Bδ) .

• Hematopoiesis: LCMT1 knockout in mice causes fetal liver hypocellularity, reduced hematopoietic stem/progenitor cells (KLS cells), and multilineage defects in repopulation assays .

• Cancer Progression: LCMT1 loss disrupts PP2A heterotrimerization, promoting phosphorylation of oncogenic substrates like MED1 and androgen receptor (AR), which drive prostate cancer therapy resistance .

Role of LCMT1 Antibodies in Research

• PP2A Complex Stability: LCMT1 antibodies (e.g., unconjugated variants) are used to assess PP2Ac methylation status, which correlates with PP2A activity and tumor suppression .

• Subcellular Localization: FITC-conjugated antibodies enable visualization of LCMT1 in organelles or cytoplasmic compartments, aiding studies on PP2A/PP4 trafficking .

Cross-Reactivity and Specificity

• Species: Human, Mouse, Rat, Monkey (predicted) .

• Control Experiments:

  • Negative Controls: Use secondary antibody alone or non-transfected cells to exclude nonspecific binding .

  • Positive Controls: Transfected cells overexpressing LCMT1 (e.g., 293T cells) .

Western Blot (WB) and Immunofluorescence (IF) Performance

FITC-conjugated variants are not explicitly validated in WB; fluorescence-based methods are preferred.

Limitations and Considerations

1. Epitope Competition: FITC conjugation may alter antibody binding affinity or epitope accessibility compared to unconjugated variants.

2. Signal Interference: Autofluorescence in tissues or overlapping fluorophores in multicolor experiments may require optimization .

3. Batch Variability: Users should validate each antibody lot for consistent performance in their experimental system.

Future Directions

The development of LCMT1 antibodies with higher sensitivity (e.g., single-chain variable fragments or nanobodies) could enhance resolution in imaging studies. Additionally, combining LCMT1 antibodies with phospho-specific probes (e.g., p-MED1 or p-AR) could elucidate PP2A-mediated signaling pathways in real-time .