Mono-Methyl-Histone H1 (Lys25) Antibody

Key Validation Metrics

• Specificity: Detects endogenous mono-methyl-K25 histone H1 without cross-reactivity to unmodified or other methylated histone H1 isoforms .

• Sensitivity: Works at dilutions of 1:500–1:1000 (Boster Bio) with optimized blocking protocols .

• Reproducibility: Boster Bio validates via WB, IHC, and ELISA using positive/negative controls . SAB Biotechnology confirms reactivity across human, mouse, and rat samples .

Example Validation Data

• Figure 1 (Boster Bio): WB of HeLa cell lysate shows a single band at ~22 kDa, consistent with HIST1H1A’s molecular weight .

• Figure 2 (SAB): Comparable results in mouse liver tissue lysates, with signal elimination after peptide competition assays .

Functional Overview

• Role: Histone H1 stabilizes higher-order chromatin structures by binding linker DNA between nucleosomes.

• Methylation at K25: Associated with transcriptional regulation and chromatin condensation during mitosis .

Gene and Protein Details

• Gene Name: HIST1H1A

• UniProt ID: Q02539

• Molecular Weight: 21,842 Da (unmodified)

• Isoforms: Includes H1.1, H1a, and H1s-1, with tissue-specific expression patterns .

Primary Use Cases

• Chromatin Dynamics: Studying methylation’s role in cell cycle progression (e.g., mitotic chromatin compaction) .

• Epigenetic Screens: Identifying K25-methylation patterns in cancer models or developmental biology.

Protocol Recommendations

• Sample Preparation: Use fresh nuclear extracts to preserve methylation states.

• Blocking: 5% non-fat milk in TBST for 1 hour at room temperature.

• Antibody Incubation: Overnight at 4°C with gentle shaking .

Limitations and Considerations

• Species Cross-Reactivity: Confirmed in human, mouse, and rat, but untested in other models .

• Post-Translational Variability: Methylation levels may vary with cell cycle stage or stress conditions.