Anti-Histone H1.5 antibody - ChIP Grade (ab18208)
Key features and details
- Rabbit polyclonal to Histone H1.5 - ChIP Grade
- Suitable for: ChIP, WB, IHC-P, ICC/IF
- Reacts with: Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-Histone H1.5 antibody - ChIP Grade
See all Histone H1.5 primary antibodies -
Description
Rabbit polyclonal to Histone H1.5 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1.5.
Read Abcam's proprietary immunogen policy -
Positive control
- Recombinant histone H1.5
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
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.
Application Abreviews Notes 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. Target
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Function
Histones H1 are necessary for the condensation of nucleosome chains into higher order structures. -
Sequence similarities
Belongs to the histone H1/H5 family.
Contains 1 H15 (linker histone H1/H5 globular) domain. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 3009 Human
- Omim: 142711 Human
- SwissProt: P16401 Human
- Unigene: 131956 Human
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Alternative names
- H1 antibody
- H1 histone family member 5 antibody
- H1.5 antibody
see all
Images
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Western blot - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)This image is courtesy of Dr Albert Jordan and Monica Sancho, Center for Genomic Regulation (CRG). Recombinant H1 isoforms courtesy of Dr Nicole Happel.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 -
ChIP - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)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.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)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.
Protocols
References (6)
ab18208 has been referenced in 6 publications.
- 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). PubMed: 31307224
- Gould TJ et al. Defining the epichromatin epitope. Nucleus 8:625-640 (2017). PubMed: 28960120
- Wang L et al. Intracellular CD24 disrupts the ARF-NPM interaction and enables mutational and viral oncogene-mediated p53 inactivation. Nat Commun 6:5909 (2015). PubMed: 25600590
- Momeni M et al. Immunohistochemical detection of promyelocytic leukemia zinc finger and histone 1.5 in uterine leiomyosarcoma and leiomyoma. Reprod Sci 21:1171-6 (2014). PubMed: 24784718
- Laserna EJ et al. Proteomic analysis of phosphorylated nuclear proteins underscores novel roles for rapid actions of retinoic acid in the regulation of mRNA splicing and translation. Mol Endocrinol 23:1799-814 (2009). PubMed: 19812389
- Sancho M et al. Depletion of human histone H1 variants uncovers specific roles in gene expression and cell growth. PLoS Genet 4:e1000227 (2008). WB ; Human . PubMed: 18927631
Images
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Western blot - Anti-Histone H1.5 antibody - ChIP Grade (ab18208) This image is courtesy of Dr Albert Jordan and Monica Sancho, Center for Genomic Regulation (CRG). Recombinant H1 isoforms courtesy of Dr Nicole Happel.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
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ChIP - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)
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.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)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.
