Product nameAnti-Histone H1.5 antibody - ChIP Grade
See all Histone H1.5 primary antibodies
DescriptionRabbit polyclonal to Histone H1.5 - ChIP Grade
Tested applicationsSuitable for: ChIP, WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1.5.
- Recombinant histone H1.5
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 ab18208 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.|
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).|
|IHC-P||1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionHistones H1 are necessary for the condensation of nucleosome chains into higher order structures.
Sequence similaritiesBelongs to the histone H1/H5 family.
Contains 1 H15 (linker histone H1/H5 globular) domain.
Cellular localizationNucleus. Chromosome.
- Information by UniProt
- H1 antibody
- H1 histone family member 5 antibody
- H1.5 antibody
All lanes : Anti-Histone H1.5 antibody - ChIP Grade (ab18208) at 0.5 µg/ml
Lane 1 : Recombinant histone H1
Lane 2 : Recombinant histone H1.1
Lane 3 : Recombinant histone H1.2
Lane 4 : Recombinant histone H1.3
Lane 5 : Recombinant histone H1.4
Lane 6 : Recombinant histone H1.5
Predicted band size: 32 kDa
Observed band size: 32 kDa
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 ab18208 (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 ab18208 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18208, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879 Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab18208 staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab18208, 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.
This product has been referenced in:
- Sang B et al. Ras-AKT signaling represses the phosphorylation of histone H1.5 at threonine 10 via GSK3 to promote the progression of glioma. Artif Cells Nanomed Biotechnol 47:2882-2890 (2019). Read more (PubMed: 31307224) »
- Gould TJ et al. Defining the epichromatin epitope. Nucleus 8:625-640 (2017). Read more (PubMed: 28960120) »