Product nameAnti-Histone H1x antibody - ChIP Grade
See all Histone H1x primary antibodies
DescriptionRabbit polyclonal to Histone H1x - ChIP Grade
SpecificityUsing western blot analysis ab31972 recognises histone H1x in HeLa nuclear extract.
Tested applicationsSuitable for: ChIP, ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Human
- HeLa nuclear extract This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab31972 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use 2 µg for 25 µg of chromatin.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).|
FunctionHistones H1 are necessary for the condensation of nucleosome chains into higher-order structures.
Tissue specificityExpressed ubiquitously.
Sequence similaritiesBelongs to the histone H1/H5 family.
Contains 1 H15 (linker histone H1/H5 globular) domain.
Cellular localizationNucleus. Chromosome.
- Information by UniProt
- H1 histone family member X antibody
- H1FX antibody
- H1X antibody
All lanes : Anti-Histone H1x antibody - ChIP Grade (ab31972) at 1 µg/ml
Lane 1 : HeLa nuclear lysate
Lane 2 : HeLa nuclear lysate with Human Histone H1x peptide (ab18052) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 35 kDa
Observed band size: 35 kDa
Additional bands at: 98 kDa (possible non-specific secondary antibody binding)
ab31972 recognises histone H1.X in HeLa nuclear extract at 35 kDa (lane 1), which is blocked using the immunizing peptide ab18052.
IHC image of Histone H1x staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31972, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab31972 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
ICC/IF image of ab31972 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31972, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
This product has been referenced in:
- Philippe TJ et al. Loss of MeCP2 in adult 5-HT neurons induces 5-HT1A autoreceptors, with opposite sex-dependent anxiety and depression phenotypes. Sci Rep 8:5788 (2018). ChIP ; Mouse . Read more (PubMed: 29636529) »
- Ito-Ishida A et al. Genome-wide distribution of linker histone H1.0 is independent of MeCP2. Nat Neurosci 21:794-798 (2018). Read more (PubMed: 29802390) »