Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830)
Key features and details
- Mouse monoclonal [mAbcam 31830] to Histone H4 - ChIP Grade
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ChIP, ICC/IF
- Reacts with: Cow, Human
- Isotype: IgG1
Related conjugates and formulations
Overview
-
Product name
Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade
See all Histone H4 primary antibodies -
Description
Mouse monoclonal [mAbcam 31830] to Histone H4 - ChIP Grade -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, WB, ChIP, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Cow, Human
Predicted to work with: Mouse, Rat -
Immunogen
-
Positive control
- This antibody gave a positive signal in the following lysates: Calf Thymus Histone Preparation Nuclear Lysate HeLa Histone Preparation Nuclear Lysate Histone H4 Recombinant Protein IHC-P: FFPE human breast fibroadenoma. ICC/IF: HeLa cell line
-
General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
-
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: 7.50
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
-
Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
mAbcam 31830 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
-
Alternative Versions
-
ChIP Related Products
-
Compatible Secondaries
-
Immunizing Peptide (Blocking)
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab31830 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 |
---|---|---|
Flow Cyt (Intra) |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
IHC-P | (1) |
Use a concentration of 0.05 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
WB | (4) |
Use a concentration of 1 µg/ml. Detects a band of approximately 13 kDa (predicted molecular weight: 14 kDa).Can be blocked with Human Histone H4 peptide (ab13843).
|
ChIP |
Use 5 µg for 25 µg of chromatin.
|
|
ICC/IF |
Use a concentration of 5 µg/ml.
|
Notes |
---|
Flow Cyt (Intra)
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
IHC-P
Use a concentration of 0.05 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 13 kDa (predicted molecular weight: 14 kDa).Can be blocked with Human Histone H4 peptide (ab13843). |
ChIP
Use 5 µg for 25 µg of chromatin. |
ICC/IF
Use a concentration of 5 µg/ml. |
Target
-
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
-
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
-
ab31830 staining Histone H4 in HeLa cells. The cells were fixed with 100% methanol (5 min), 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 ab31830 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
-
All lanes : Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg/ml
Lane 2 : HeLa Nuclear Lysate (Triton enriched) at 10 µg/ml
Lane 3 : Histone H4 Recombinant Protein at 0.1 µg/ml
Lane 4 : Histone H3.1 Recombinant Protein at 0.1 µg/ml
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Additional bands at: 13 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 seconds -
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, 5µg of ab31830 (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.
-
Overlay histogram showing HeLa cells stained with ab31830 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab31830, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
-
All lanes : Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 2 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg
Lane 3 : Histone H4 Recombinant Protein at 0.1 µg
Lane 4 : Histone H3.1 Recombinant Protein at 0.1 µg
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted? -
All lanes : Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830) at 5 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 2 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg
Lane 3 : Histone H4 Recombinant Protein at 0.1 µg
Lane 4 : Histone H3.1 Recombinant Protein at 0.1 µg
Lane 5 : Histone H2A Recombinant Protein at 0.1 µg
Lane 6 : Histone H2B Recombinant Protein at 0.1 µg
Lane 7 : Histone H1 Recombinant Protein at 0.1 µg
Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Lane 9 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Lane 10 : Histone H4 Recombinant Protein at 0.1 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Lane 11 : Histone H3.1 Recombinant Protein at 0.1 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Lane 12 : Histone H2A Recombinant Protein at 0.1 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Lane 13 : Histone H2B Recombinant Protein at 0.1 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Lane 14 : Histone H1 Recombinant Protein at 0.1 µg withHuman Histone H4 peptide (ab13843) at 5 µg/ml
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes -
IHC image of Histone H4 staining in human breast fibroadenoma 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 ab31380, 0.1µg/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.
Datasheets and documents
-
Datasheet download
References (51)
ab31830 has been referenced in 51 publications.
- Samejima I et al. Mapping the invisible chromatin transactions of prophase chromosome remodeling. Mol Cell 82:696-708.e4 (2022). PubMed: 35090599
- Witus SR et al. BRCA1/BARD1 site-specific ubiquitylation of nucleosomal H2A is directed by BARD1. Nat Struct Mol Biol 28:268-277 (2021). PubMed: 33589814
- Fan Y et al. LRR1-mediated replisome disassembly promotes DNA replication by recycling replisome components. J Cell Biol 220:N/A (2021). PubMed: 34037657
- Wimmers F et al. The single-cell epigenomic and transcriptional landscape of immunity to influenza vaccination. Cell 184:3915-3935.e21 (2021). PubMed: 34174187
- Campiani G et al. Harnessing the Role of HDAC6 in Idiopathic Pulmonary Fibrosis: Design, Synthesis, Structural Analysis, and Biological Evaluation of Potent Inhibitors. J Med Chem 64:9960-9988 (2021). PubMed: 34251197