Overview

  • Product name
  • Description
    Rabbit polyclonal to Histone H4
  • Host species
    Rabbit
  • Specificity
    We recommend ab16483 as a suitable alternative to ab10156 as it was made using the same immunogen.
  • Tested applications
    Suitable for: WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Cow, Human, Saccharomyces cerevisiae
  • Immunogen

    Recombinant full length protein corresponding to Saccharomyces cerevisiae Histone H4 aa 1 to the C-terminus.
    Database link: P02309

  • Positive control
    • Calf thymus histone lysate and Hela histone lysate.

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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG

Applications

Our Abpromise guarantee covers the use of ab16483 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 Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).
Flow Cyt Use at an assay dependent concentration. PubMed: 20709786

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

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
    modifications
    Acetylation 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
  • Alternative names
    • dJ160A22.1 antibody
    • dJ160A22.2 antibody
    • dJ221C16.1 antibody
    • dJ221C16.9 antibody
    • FO108 antibody
    • H4 antibody
    • H4.k antibody
    • H4/a antibody
    • H4/b antibody
    • H4/c antibody
    • H4/d antibody
    • H4/e antibody
    • H4/g antibody
    • H4/h antibody
    • H4/I antibody
    • H4/j antibody
    • H4/k antibody
    • H4/m antibody
    • H4/n antibody
    • H4/p antibody
    • H4_HUMAN antibody
    • H4F2 antibody
    • H4F2iii antibody
    • H4F2iv antibody
    • H4FA antibody
    • H4FB antibody
    • H4FC antibody
    • H4FD antibody
    • H4FE antibody
    • H4FG antibody
    • H4FH antibody
    • H4FI antibody
    • H4FJ antibody
    • H4FK antibody
    • H4FM antibody
    • H4FN antibody
    • H4M antibody
    • HIST1H4A antibody
    • HIST1H4B antibody
    • HIST1H4C antibody
    • HIST1H4D antibody
    • HIST1H4E antibody
    • HIST1H4F antibody
    • HIST1H4H antibody
    • HIST1H4I antibody
    • HIST1H4J antibody
    • HIST1H4K antibody
    • HIST1H4L antibody
    • HIST2H4 antibody
    • HIST2H4A antibody
    • Hist4h4 antibody
    • Histone 1 H4a antibody
    • Histone 1 H4b antibody
    • Histone 1 H4c antibody
    • Histone 1 H4d antibody
    • Histone 1 H4e antibody
    • Histone 1 H4f antibody
    • Histone 1 H4h antibody
    • Histone 1 H4i antibody
    • Histone 1 H4j antibody
    • Histone 1 H4k antibody
    • Histone 1 H4l antibody
    • Histone 2 H4a antibody
    • histone 4 H4 antibody
    • Histone H4 antibody
    • MGC24116 antibody
    see all

Images

  • All lanes : Anti-Histone H4 antibody (ab16483) at 1 µg/ml

    Lane 1 : Calf Thymus Histone lysate at 0.5 µg
    Lane 2 : HeLa Histone Lysate at 2.5 µg
    Lane 3 : Recombinant Histone H4 at 1 µg

    Secondary
    All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Predicted band size: 11 kDa
    Observed band size: 14 kDa
    why is the actual band size different from the predicted?



    ab16483 detects a band of approximately 14 kDa in Calf thymus histone prep, HeLa histone prep and recombinant Histone H4. This is slightly larger than the predicted molecular weight according to Swissprot (11 kDa), however we are confident that ab16483 is recognising Histone H4.

References

This product has been referenced in:
See all 9 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for contacting Abcam.
We have not tested either of these antibodies in ChIP, however either may be suitable but I did find ab7311, which we do guarantee to work in ChIP.
If you would like to try either ab10158 or ab16483 in ChIP, then either would qualify for our testing discoutn program, the link below will provide more details about the testing discount program:
https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount
If there is anything else I can help you with, please let me know.

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Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 88739 DESCRIPTION OF THE PROBLEM No band after the western blot hyb with this antibody following your recommended concentration (1?g/ml). Simultaneously, I performed a Western blot of with other Abcam antibodies and got a correct and clear band (unmodified H3 and several antibodies recognizing acetylated and methylated forms of histones. SAMPLE Purified Histone solution+DNA (I have performed a binding assay in which I bound purified histones+DNA). As I stated above, I already did it with other antibodies (I used the same conditions) and worked well. PRIMARY ANTIBODY Abcam 16483, 1?g/ml in 10 ml of TBST 5%milk for 2 or 4 hours (I did it 2 times) and wash with TBST (3x15') DETECTION METHOD ECL plus POSITIVE AND NEGATIVE CONTROLS USED positive control: purified histone solution (previously checked with other antibodies that worked well, i.e. clear signal) ANTIBODY STORAGE CONDITIONS Stored at +4. SAMPLE PREPARATION Standard and published histone sample preparation (+ standard Laemmli loading buffer... Exactly same conditions that worked well for other antibodies... AMOUNT OF PROTEIN LOADED as recommended, 1?g/ml ELECTROPHORESIS/GEL CONDITIONS gradient 10-20% TRANSFER AND BLOCKING CONDITIONS 4 hours transfer in a glycine-Tris-methanol buffer, checked by red punceau (the histones were there as determined by a positive control) and used TBST 5% milk blocking buffer. Same conditions that worked well for other antibodies (run in parallel) SECONDARY ANTIBODY ECL Antirabbit IgG, dilution 1/10000 and 1/1000 in 10 ml of TBST 5%milk for 2 hours (I did it 2 times) and wash with TBST (3x15'). All the conditions worked well for other antibodies (run in parallel). HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I did longer incubations and increased amount of secondary antibody ADDITIONAL NOTES After the transfer, the histones were there (determiined by red punceau staining). Simultaneously, I performed a Western blot of with other Abcam antibodies and got a correct and clear band (unmodified H3 and several antibodies recognizing acetylated and methylated forms of histones.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionaire and you are using an approach that I would certainly recommend. I am certainly prepared to offer you credit against your purchase provided that you purchased this antibody within the past 90 days. If this is the case please email me details of your order including the number and date.

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