Recombinant Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (ab45166)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1002Y] to Histone H4 (acetyl K8) - ChIP Grade
- Suitable for: Flow Cyt (Intra), ChIP, ChIP-sequencing, WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade
See all Histone H4 primary antibodies -
Description
Rabbit monoclonal [EP1002Y] to Histone H4 (acetyl K8) - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ChIP, ChIP-sequencing, WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Histone H4 aa 1-100 (N terminal) (acetyl K8). The exact sequence is proprietary.
Database link: P62805 -
Positive control
- WB: HeLa whole cell lysate +TSA, C6 cell lysate, C6 cell + TSA lysate, NIH/3T3 +TSA whole cell lysate. IHC-P: Human normal colon FFPE tissue sections, mouse kidney paraffin-embedded tissue sections, rat kidney paraffin-embedded tissue sections. ICC/IF: C6 + TSA lysates. ChIP: Chromatin prepared from HeLa cells ChiP-Seq: HeLa Cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.17% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1002Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab45166 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ChIP | (1) |
Use 2 µg for 25 µg of chromatin.
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ChIP-sequencing |
Use 4µg for 107 cells.
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WB |
1/5000 - 1/10000. Predicted molecular weight: 11 kDa.
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IHC-P |
1/250 - 1/2500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/150 - 1/500.
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IP |
1/20 - 1/50.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ChIP
Use 2 µg for 25 µg of chromatin. |
ChIP-sequencing
Use 4µg for 107 cells. |
WB
1/5000 - 1/10000. Predicted molecular weight: 11 kDa. |
IHC-P
1/250 - 1/2500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/150 - 1/500. |
IP
1/20 - 1/50. |
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 H4 family. -
Post-translational
modificationsAcetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
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. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
Sumoylated, which is associated with transcriptional repression. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 121504 Human
- Entrez Gene: 554313 Human
- Entrez Gene: 8294 Human
- Entrez Gene: 8359 Human
- Entrez Gene: 8360 Human
- Entrez Gene: 8361 Human
- Entrez Gene: 8362 Human
- Entrez Gene: 8363 Human
see all -
Alternative names
- dJ160A22.1 antibody
- dJ160A22.2 antibody
- dJ221C16.1 antibody
see all
Images
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Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 4 µg of ab45166 [EP1002Y]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
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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 ab45166 (red), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
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All lanes : Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (ab45166) at 1/5000 dilution
Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma) treated with Trichostatin A whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking and diluting buffer 5% NFDM/TBST -
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified ab45166 at 1/20 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized C6 (rat glioma) cells (non-treated-top panels) and (C6 + TSA(500ng/ml, 4hr)-middle panels) with purified ab45166 at dilution of 1/150. The secondary antibody used was Alexa Fluor® 488; goat anti-rabbit IgG (ab150077) at a dilution of 1/1000. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/1000) was used to stain tubulin along with ab150120 (AlexaFluor®594 goat anti-mouse secondary, 1/1000) shown in the top right and middle right hand panels. The negative controls are shown in the bottom two panels- for negative control 1 rabbit primary antibody and anti-mouse secondary antibody (ab150120) was used. For negative control 2 mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) was used.
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Immunohistochemical staining of paraffin-embedded mouse kidney sections labelling Histone H4 (acetyl K8) with purified ab45166 at dilution of 1:2500. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
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ab45166 (purified) at 1/20 immunoprecipitating Histone H4 (acetyl K8) in HeLa treated with Trichostatin A whole cell lysate.
Lane 1 (input): HeLa treated with Trichostatin A whole cell lysate (10µg)
Lane 2 (+): ab45166 + HeLa treated with Trichostatin A whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab45166 in HeLa treated with Trichostatin A whole cell lysate.
For western blotting, ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/10000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Lanes 1-2 : Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (ab45166) at 1/20 dilution
Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) at 1/20 dilution
Lane 1 : HeLa (human cervix adenocarcinoma) treated with Trichostatin A whole cell lysate at 10 µg
Lanes 2-3 : HeLa (human cervix adenocarcinoma) treated with Trichostatin A whole cell lysate
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution
Observed band size: 11 kDa why is the actual band size different from the predicted? -
Immunohistochemical staining of paraffin-embedded rat kidney sections labelling Histone H4 (acetyl K8) with purified ab45166 at dilution of 1:2500. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
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All lanes : Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (ab45166) at 1/5000 dilution
Lane 1 : Untreated C6 (rat glioma) whole cell lysate
Lane 2 : C6 (rat glioma) treated with Trichostatin A whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking and diluting buffer 5% NFDM/TBST -
Immunohistochemical analysis of formalin fixed paraffin embedded human colon tissue sections labelling Histone H4 (acetyl K8) with unpurified ab45166 at dilution of 1/200.
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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All lanes : Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (ab45166) at 1/5000 dilution
Lane 1 : Untreated NIH/3T3 (mouse embryo) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryo) treated with Trichostatin A whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking and diluting buffer 5% NFDM/TBST -
Immunohistochemical staining of paraffin-embedded human colon sections labelling Histone H4 (acetyl K8) with purified ab45166 at dilution of 1:2500. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
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 (21)
ab45166 has been referenced in 21 publications.
- Van HT et al. Methyl-lysine readers PHF20 and PHF20L1 define two distinct gene expression-regulating NSL complexes. J Biol Chem 298:101588 (2022). PubMed: 35033534
- Sun YC et al. Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium. Invest Ophthalmol Vis Sci 63:31 (2022). PubMed: 35212722
- Srivastava RK et al. Combined inhibition of BET bromodomain and mTORC1/2 provides therapeutic advantage for rhabdomyosarcoma by switching cell death mechanism. Mol Carcinog 61:737-751 (2022). PubMed: 35472745
- Chomiak AA et al. Nde1 is required for heterochromatin compaction and stability in neocortical neurons. iScience 25:104354 (2022). PubMed: 35601919
- Liu X et al. LINC00839 promotes colorectal cancer progression by recruiting RUVBL1/Tip60 complexes to activate NRF1. EMBO Rep 23:e54128 (2022). PubMed: 35876654