Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684)
Key features and details
- Rabbit polyclonal to Histone H3 (di methyl K27) - ChIP Grade
- Suitable for: WB, IHC-P, ICC/IF, ChIP
- Reacts with: Mouse, Cow, Human, Caenorhabditis elegans
- Isotype: IgG
Overview
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Product name
Anti-Histone H3 (di methyl K27) antibody - ChIP Grade
See all Histone H3 primary antibodies -
Description
Rabbit polyclonal to Histone H3 (di methyl K27) - ChIP Grade -
Host species
Rabbit -
Specificity
This antibody is specific for Histone H3 di-methylated at residue K27. It does not recognise the mono- or tri-methylated K27 residue or the mono-/di-/tri-methylated H3 K9 residue. -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, ChIPmore details -
Species reactivity
Reacts with: Mouse, Cow, Human, Caenorhabditis elegans
Predicted to work with: Saccharomyces cerevisiae, Drosophila melanogaster, Plants, Cyanidioschyzon merolae
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Immunogen
Synthetic peptide corresponding to Histone H3 aa 1-100 (N terminal) (di methyl K27) conjugated to keyhole limpet haemocyanin.
(Peptide available asab1781) -
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Control Peptide
- Human Histone H3 (mono methyl K4) peptide (ab1340)
- Human Histone H3 (tri methyl K4) peptide (ab1342)
- Human Histone H3 (mono methyl K9) peptide (ab1771)
- Human Histone H3 (di methyl K9) peptide (ab1772)
- Human Histone H3 (mono methyl K27) peptide (ab1780)
- Human Histone H3 (tri methyl K27) peptide (ab1782)
- Human Histone H3 (unmodified ) peptide (ab2623)
- Human Histone H3 (di methyl K4) peptide (ab7768)
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab24684 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 |
|---|---|---|
| WB | (8) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15.2 kDa).Can be blocked with Human Histone H3 (di methyl K27) peptide (ab1781).
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| IHC-P | (8) |
Use a concentration of 1 µg/ml.
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| ICC/IF | (5) |
Use a concentration of 1 µg/ml.
Suitable for cells that have been fixed with 100% Methanol (5 mins) or 4% PFA (10 mins). |
| ChIP |
Use 2-25 µg for µg of chromatin.
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| Notes |
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WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15.2 kDa).Can be blocked with Human Histone H3 (di methyl K27) peptide (ab1781). |
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IHC-P
Use a concentration of 1 µg/ml. |
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ICC/IF
Use a concentration of 1 µg/ml. Suitable for cells that have been fixed with 100% Methanol (5 mins) or 4% PFA (10 mins). |
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ChIP
Use 2-25 µg for µg of chromatin. |
Target
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Function
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. -
Sequence similarities
Belongs to the histone H3 family. -
Developmental stage
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. -
Post-translational
modificationsAcetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
- Entrez Gene: 8356 Human
- Entrez Gene: 8357 Human
see all -
Alternative names
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684)
ab24684 staining Histone H3 (di methyl K27) in HeLa cells. The cells were fixed with 100% Methanol (5min), permeabilized with 0.1%PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab24684 at 1μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684)IHC image of Histone H3 (di methyl K27) staining in Human normal 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 ab24684, 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.
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Western blot - Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684)All lanes : Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684) at 1 µg/ml
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with Human Histone H3 peptide (ab17163) at 0.5 µg/ml
Lane 3 : Calf thymus histone lysate withHuman Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
Lane 4 : Calf thymus histone lysate withHuman Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
Lane 5 : Calf thymus histone lysate withHuman Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
Lane 6 : Calf thymus histone lysate withHuman Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
Lane 7 : Calf thymus histone lysate withHuman Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lane 8 : Calf thymus histone lysate withHuman Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
Lane 9 : Calf thymus histone lysate withHuman Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/ml
Lane 10 : Calf thymus histone lysate withHuman Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/ml
Lane 11 : Calf thymus histone lysate withHuman Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg/ml
Secondary
All lanes : Goat polyclonal to Rabbit IgG H&L (HRP) at 1/5000 dilution
Predicted band size: 15.2 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsThe peptide competition assay shows that this antibody is very specific for Histone H3 di-methylated at K27. It does not recognise Histone H3 mono
, di , or tri methylated at K4 , Histone H3 mono , di , or tri methylated at K9 , or Histone H3 mono , or tri methylated at K27. -
ChIP - Anti-Histone H3 (di methyl K27) antibody - ChIP Grade (ab24684)Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab24684 (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 H3 (di methyl K27) antibody - ChIP Grade (ab24684)This image is courtesy of an anonymous abreview.Immunocytochemistry/ Immunofluorescence analysis of IMR32, Human Neuroblatoma cells labeling Histone H3 (di methyl K27) with ab24684 at 1/100 dilution. The cells were fixed with paraformaldehyde and permeabilized with 0.3% triton X 100. The cells were blocked with 1% BSA for 1 hour at 25°C, followed by incubation with primary antibody in 1% BSA containing TBST for 16 hours at 4°C. A Goat Anti Rabbit-IgG Cy3® was used as the secondary antibody at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (93)
ab24684 has been referenced in 93 publications.
- Liu Q et al. Histone demethylase PHF8 drives neuroendocrine prostate cancer progression by epigenetically upregulating FOXA2. J Pathol 253:106-118 (2021). PubMed: 33009820
- Li TY et al. The transcriptional coactivator CBP/p300 is an evolutionarily conserved node that promotes longevity in response to mitochondrial stress. Nat Aging 1:165-178 (2021). PubMed: 33718883
- Liu J et al. Maternal Diabetes-Induced Suppression of Oxytocin Receptor Contributes to Social Deficits in Offspring. Front Neurosci 15:634781 (2021). PubMed: 33633538
- Xie W et al. Pterostilbene accelerates wound healing by modulating diabetes-induced estrogen receptor ß suppression in hematopoietic stem cells. Burns Trauma 9:tkaa045 (2021). PubMed: 33654697
- Yang Y et al. Modulators of histone demethylase JMJD1C selectively target leukemic stem cells. FEBS Open Bio 11:265-277 (2021). PubMed: 33289299