Product nameAnti-Histone H4 antibody [IGX4696H] - ChIP Grade
See all Histone H4 primary antibodies
DescriptionHuman monoclonal [IGX4696H] to Histone H4 - ChIP Grade
Tested applicationsSuitable for: WB, ChIPmore details
Species reactivityReacts with: Mouse, Human
Database link: P62805
- WB: CTH, HeLa whole cell lysate, HeLa nuclear lysate, NIH3T3 whole cell lysate, NIH3T3 nuclear lysate and Histone H4B Recombinant Protein. ChIP: HeLa and NIH3T3.
This product was made using synthetic libraries and phage display technology.
This antibody is a recombinant antibody.
Human monoclonal antibody.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium azide
Constituents: 1% BSA, PBS
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab213291 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.2 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).|
|ChIP||Use 2 µg for 25 µg of chromatin.|
FunctionCore 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 similaritiesBelongs to the histone H4 family.
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 localizationNucleus. Chromosome.
- Information by UniProt
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All lanes : Anti-Histone H4 antibody [IGX4696H] - ChIP Grade (ab213291) at 0.2 µg/ml
Lane 1 : CTH (Calf Thymus Histone) at 0.5 µg
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear Lysate at 10 µg
Lane 6 : Histone H2A Recombinant Protein at 0.1 µg
Lane 7 : Histone H4 Recombinant Protein at 0.1 µg
All lanes : HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab213291 overnight at 4°C. Antibody binding was detected using an anti-human antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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 ab213291 (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). Primers and probes are located in the first kb of the transcribed region.
Chromatin was prepared from NIH3T3 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 ab213291 (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 (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.
ab213291 has not yet been referenced specifically in any publications.