HIST1H1C (Ab-33) Antibody
Description
Technical Notes
-
Epitope Specificity: The antibody binds to a conserved region of HIST1H1C, enabling cross-reactivity with homologous sequences in multiple species (e.g., mouse, rat, cow) .
-
Validation: Demonstrated efficacy in Western Blot using cell lysates as positive controls .
-
Storage: Aliquots stored at -20°C to prevent degradation; repeated freeze-thaw cycles should be avoided .
Applications in Research
While direct studies using the HIST1H1C (Ab-33) antibody are not explicitly detailed in the provided literature, its utility aligns with broader research on HIST1H1C’s roles:
Chromatin Structure and Gene Regulation
HIST1H1C is implicated in chromatin compaction and the regulation of transcriptional programs. Antibodies targeting this histone variant are critical for studying:
-
Nucleosome dynamics: H1 variants influence linker DNA accessibility and higher-order chromatin organization .
-
Epigenetic modifiers: HIST1H1C interacts with histone deacetylases (e.g., HDAC1) to modulate acetylation states (e.g., H4K16 deacetylation) .
Disease Pathologies
Research on HIST1H1C highlights its involvement in:
-
Viral replication: HIST1H1C inhibits influenza A virus by upregulating interferon-β (IFN-β) and enhancing IRF3 nuclear translocation .
-
Diabetic retinopathy: Overexpression promotes autophagy, inflammation, and neuronal loss, suggesting therapeutic targeting .
-
Hepatocarcinogenesis: Elevated HIST1H1C levels correlate with STAT3 activation and tumor progression in HCC .
Limitations and Considerations
-
Cross-reactivity: Predicted reactivity with non-human species (e.g., cow, dog) requires experimental validation .
-
Specificity: No data on cross-reactivity with other histone H1 variants (e.g., H1.3, H1.4) are provided in available sources.
-
Experimental Design: Optimal dilutions and protocols must be optimized for individual assays (e.g., ChIP, IHC) .
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research has identified a network of E2F target genes susceptible to the regulatory influence of H1.2. H1.2 enhances the global association of pRb with chromatin, strengthens transcriptional repression by pRb, and facilitates pRb-dependent cell-cycle arrest. PMID: 28614707
- BRG1 participates in gene repression by interacting with H1.2, promoting its deposition and stabilizing nucleosome positioning around the transcription start site. PMID: 27390128
- Studies have observed histones H1.2 and H1.4 in MDA-MB-231 metastatic breast cancer cells. Phosphorylation at S173 of histone H1.2 and S172, S187, T18, T146, and T154 of H1.4 significantly increases during the M phase, indicating a cell cycle-dependent nature. Additionally, the study reports the observation of the H1.2 SNP variant A18V in MCF-10A cells. PMID: 26209608
- Integration with apoptotic intermediates (via C-terminal tail interactions) may represent a more generalized function of linker histone isoforms in apoptotic cascades. PMID: 24525734
- Post-translational modifications of H1.2-T165 are dispensable for chromatin binding and cell proliferation, while modifications of H1.4-K26 are essential for proper cell cycle progression. PMID: 24873882
- H1.2 interacts with Cul4A and PAF1 to activate developmental regulatory genes. PMID: 24360965
- H1.2 is less abundant than other histone H1 variants at the transcription start sites of inactive genes. Promoters enriched in H1.2 differ from those enriched in other histone H1 variants and tend to be repressed. PMID: 24476918
- Mutations in linker histone genes HIST1H1 B, C, D, and E; OCT2 (POU2F2); IRF8; and ARID1A are associated with the pathogenesis of follicular lymphoma. PMID: 24435047
- Data suggests that the p53 acetylation-H1.2 phosphorylation cascade serves as a unique mechanism for triggering p53-dependent DNA damage response pathways. PMID: 22249259
- Research has confirmed N-terminal acetylation on all isoforms, plus a single internal acetylation site. Phosphorylation sites were located on peptides containing the cyclin-dependent kinase (CDK) consensus motif. PMID: 15595731
- The binding of histone H1 to a general amyloid-like motif suggests that histone H1 may play a crucial role in diseases associated with amyloid-like fibrils. PMID: 16854430
- Histone H1.2 translocates from the nucleus to the mitochondria after treatment with bleomycin and co-localizes with Bak in mitochondria. PMID: 17879944
- Research has shown that the recruitment of YB1, PURalpha, and H1.2 to the p53 target gene Bax is necessary for the repression of p53-induced transcription. PMID: 18258596
Q&A
What are the primary experimental applications of HIST1H1C (Ab-33) antibody in molecular biology studies?
This polyclonal antibody is validated for three core applications:
-
Western Blotting (1:100-1:1000 dilution) to detect histone H1.2 variants in nuclear extracts
-
Immunohistochemistry (1:20-1:200 dilution) for chromatin structure analysis in retinal tissues
-
ELISA quantification of histone H1c epigenetic modifications under diabetic conditions
Key methodological considerations include:
-
Nuclear fraction isolation requirement for WB applications due to HIST1H1C's chromatin-binding nature
-
Antigen retrieval optimization for IHC in formaldehyde-fixed neural tissues
-
Parallel H3/H4 histone controls to confirm assay specificity
How should researchers validate HIST1H1C antibody specificity in epigenetic studies?
A three-tier validation protocol is recommended:
Recent studies show 92% concordance between HIST1H1C (Ab-33) and mass spectrometry results in diabetic retinopathy models when using proper chromatin shearing protocols .
How does HIST1H1C regulate autophagy pathways in diabetic retinopathy pathogenesis?
The histone exhibits dual regulatory mechanisms:
Mechanism 1: Upregulates SIRT1/HDAC1 complex (2.3-fold increase), reducing H4K16 acetylation (−40%) and activating ATG genes (Becn1, Map1lc3b)
Mechanism 2: Promotes LC3B-II conversion (1.8× baseline) through SQSTM1/p62 degradation (−55% protein levels)
Critical experimental parameters:
-
Glucose concentration threshold: 25mM induces maximal HIST1H1C effect in Müller cells
-
Temporal dynamics: Autophagy peaks at 72hrs post-HIST1H1C transfection
What experimental strategies resolve cytoplasmic/nuclear localization discrepancies of HIST1H1C?
Addressing the observed 34% cytoplasmic staining in diabetic retinas vs <5% in vitro :
Solution 1: Implement in situ proximity ligation assays to differentiate true translocation vs extraction artifacts
Solution 2: Use compartment-specific markers (Lamin B1 nuclear, GAPDH cytoplasmic) in fractionation protocols
Solution 3: Employ high-resolution STED microscopy (30nm resolution) for subnuclear localization analysis
Recent findings suggest caspase-mediated cleavage (at DXXD motifs) enables mitochondrial translocation under oxidative stress, explaining in vivo vs in vitro differences .
How to design CRISPR studies investigating HIST1H1C-viral protein interactions?
The influenza study provides a validated framework:
| Component | Specification |
|---|---|
| Guide RNA | 5'-AACCAATGTCACCGGCGCCGGCC-3' (Exon 2 targeting) |
| Validation | Sanger sequencing + MS-based histone profiling |
| Phenotypic Assays | Viral NP mRNA quantification (ddPCR recommended) |
Critical parameters:
-
Maintain <0.5% FBS during selection to prevent compensatory histone expression
-
Use H1C-KO A549 cells + WT rescue controls (≥3 biological replicates)
-
Monitor interferon-β levels (ELISA sensitivity: 15pg/mL required)
Methodological Comparison Table
Data from demonstrate technique-dependent variability in HIST1H1C detection efficiency, with IHC showing 22% greater sensitivity than WB in neural tissues.
Contradiction Analysis Framework
Conflict: Reported pro-autophagic vs anti-viral effects
Resolution Strategy:
-
Contextualize cellular stress conditions (diabetic vs infected states)
-
Map interaction partners:
-
Quantify post-translational modifications:
Experimental validation should combine SILAC-based proteomics (2× histone labeling) with site-directed mutagenesis (K34A/T146A variants).
