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Product name
Anti-Histone H1 (tri methyl K25) antibody
Description
Rabbit polyclonal to Histone H1 (tri methyl K25)
Specificity
ab17347 gives a positive result against the immunizing peptide and a negative result against the corresponding unmodified peptide in ELISA analysis. ab17347 recognises histone H1 tri methyl K25 in HeLa nuclear and whole cell lysates at 35 kDa. The signal is efficiently blocked using the immunzing histone H1 tri methyl K25 peptide but not the corrseponding unmodified peptide or histone H1 di methyl K25 peptide. ab17347 also appears to recognise other bands between 34 kDa and 37 kDa, which are attributed to the antibody recognising other isoforms of histone H1. ab17347 also appears to recognise methylated histone H3 at 17 kDa.
Tested applications
IHC-FoFr, WB, ICC/IF, IHC-Pmore details
Cross reactivity
Reacts with
Cow, Human
Predicted to work with
Non Human Primates
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1, tri methylated at K25.
Positive control
Hela whole cell lysate
General notes
HeLa nuclear lysate
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> Histones >> H1 >> Methylated
Applications
Show applications key
Our Abpromise guarantee covers the use of ab17347 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-FoFr
IHC-FoFr: 1/200.IHC-FoFr: 1/200.
WB: Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).Can be blocked with Histone H1 peptide - tri methyl K25 (ab17587).
ICC/IF: Use a concentration of 1 µg/ml.
IHC-P: 1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Target
Relevance
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Linker histones are involved in the formation of higher order structure in chromatin and the maintenance of overall chromatin compaction. Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fibre is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Methylation of position-specific lysine residues in histone N termini is a central modification for regulating epigenetic transitions in chromatin. Each methylatable lysine residue can exist in a mono-, di-, or trimethylated state. Arginine resdiues can also by mono or di methylated.
Cellular localization
Nuclear
Alternative names
- H1 antibody
- H1.4 antibody
- Histone H1.4 antibody
Database links
- Entrez Gene: 3005 Human
- Omim: 142708 Human
- SwissProt: P10412 Human
- Unigene: 226117 Human
Anti-Histone H1 (tri methyl K25) antibody images:
Western blot - Histone H1 (tri methyl K25) antibody (ab17347)
All lanes : Anti-Histone H1 (tri methyl K25) antibody (ab17347) at 1 µg/ml
Lane 1 : HeLa nuclear lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : HeLa nuclear lysate with
Lane 4 : HeLa whole cell lysate with
Lane 5 : HeLa nuclear lysate with
Lane 6 : HeLa whole cell lysate with
Lane 7 : HeLa nuclear lysate with Histone H1 peptide - di methyl K25 (ab21998) at 1 µg/ml
Lane 8 : HeLa whole cell lysate with Histone H1 peptide - di methyl K25 (ab21998) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution at 1/10000 dilution
Predicted band size : 35 kDa
Observed band size : 35 kDa
Additional bands at : 17 kDa (possible cross reactivity).
ab17347 recognises histone H1 tri methyl K25 in HeLa nuclear (lane1) and whole cell (lane2) lysates at 35 kDa. The signal is efficiently blocked using the immunizing histone H1 tri methyl K25 peptide (lanes3-4) but not the corrseponding unmodified peptide (lanes5-6) or histone H1 di methyl K25 peptide (lanes7-8).
ab17347 also appears to recognise other bands between 34 kDa and 37 kDa, which are attributed to the antibody recognising other isoforms of histone H1.
The strong signal at 17 kDa is attributed methylated histone H3.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H1 (tri methyl K25) antibody (ab17347)
Image courtesy of Human Protein Atlas
ab17347 staining histone H1 tri methyl K25 in female cerebellum, showing a distinct and strong staining pattern at cells in the granular and molecular layers. Paraffin embedded human skin tissue was incubated with ab17347(1/300 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab17347 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Immunocytochemistry/ Immunofluorescence - Histone H1 (tri methyl K25) antibody (ab17347)
ICC/IF image of ab17347 stained human HepG2 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab17347, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to 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 antibody also gave a positive IF result in HeLa, HEK 293, MCF7 cells.
References for Anti-Histone H1 (tri methyl K25) antibody (ab17347)
This product has been referenced in:
- Agger K et al. UTX and JMJD3 are histone H3K27 demethylases involved in HOX gene regulation and development. Nature 449:731-4 (2007). ICC/IF; Human.Read more (PubMed: 17713478) »
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"