Overview

  • Product name
    Anti-Histone H4 antibody - ChIP Grade
    See all Histone H4 primary antibodies
  • Description
    Rabbit polyclonal to Histone H4 - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    Histone H4
  • Tested applications
    Suitable for: IHC-P, ICC/IF, ChIP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Saccharomyces cerevisiae
    Predicted to work with: Mammals
  • Immunogen

    Synthetic peptide corresponding to Human Histone H4 aa 1-100 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P62805
    (Peptide available as ab21044)

  • Positive control
    • Calf Thymus Histone Preparation; Hela whole cell extract

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
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7311 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/200.
ChIP Use 2 µg for 25 µg of chromatin.
WB 1/500 - 1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).

Target

Images

  • 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 ab7311 (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.

  • IHC image of Histone H4 staining in human breast carcinoma FFPE section, performed on a 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 ab7311, 1µg/ml, for 8 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.
  • All lanes : Anti-Histone H4 antibody - ChIP Grade (ab7311) at 1/500 dilution

    Lane 1 : Calf thymus histone extract
    Lane 2 : Calf thymus histone extract with Human Histone H4 peptide (ab21044) at 1 µg/ml
    Lane 3 : Calf thymus histone extract with Histone H4 peptide - acetyl K16 (ab22376) at 1 µg/ml
    Lane 4 : Calf thymus histone extract with Human Histone H4 (acetyl K20) peptide (ab20632) at 1 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : Alexa-Fluor Goat anti-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?

  • SKN-SH cells were fixed in 4% paraformaldehyde for 10 mins, permeabilized in PBS-0.5% Triton X-100 for 5 mins and incubated for 30 minutes with ab7311 (1/200 dilution). The slides were rinsed once in PBS-Triton (0.1%), twice in PBS then incubated with the secondary antibody for 30 mins. The DNA is stained with DAPI (blue). Clear nuclear staining with ab7311 can be seen (green).

  • Anti-Histone H4 antibody - ChIP Grade (ab7311) at 1/500 dilution + Human Bone Whole Tissue Lysate at 25 µg with BSA, 1 Hour, 23°C at 5 %

    Developed using the ECL technique.

    Predicted band size: 11 kDa


    Exposure time: 1 minute

    See Abreview

References

This product has been referenced in:
  • Young CP  et al. Distinct histone methylation and transcription profiles are established during the development of cellular quiescence in yeast. BMC Genomics 18:107 (2017). WB ; Saccharomyces cerevisiae . Read more (PubMed: 28122508) »
  • Johnson WL  et al. RNA-dependent stabilization of SUV39H1 at constitutive heterochromatin. Elife 6:N/A (2017). Read more (PubMed: 28760200) »
See all 51 Publications for this product

Customer reviews and Q&As

1-10 of 44 Abreviews or Q&A

Application
ChIP
Sample
Mouse Tissue lysate - nuclear (Hippocampus)
Negative control
Normal rabbit IgG
Specification
Hippocampus
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde in PBS
Positive control
One active gene (ActB) and one repressed gene (HoxA1). The ChIP show higher levels of H4 in the repressed gene.

Abcam user community

Verified customer

Submitted Sep 06 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa Kyoto)
Permeabilization
No
Specification
HeLa Kyoto
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Methanol

Dr. Andrew Bowman

Verified customer

Submitted Apr 21 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (multiple myeloma cell)
Loading amount
50 µg
Specification
multiple myeloma cell
Gel Running Conditions
Reduced Denaturing (10-20% tris glycine gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Nov 21 2012

Answer

Thanks in advance for filling out the questionnaire below for each antibody. The answers will help me to assist you most efficiently.

1) Abcam product code ab



2) Abcam order reference number or product batch number



3) Description of the problem



4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…):

Lysis buffer

Protease inhibitors:

Phosphatase inhibitors

Reducing agent

Boiling for ≥5 min? yes/no

Protein loaded ug/lane or cells/lane

Positive control

Negative control



5) Percentage of gel

Type of membrane

Protein transfer verified

Blocking agent and concentration

Blocking time

Blocking temperature



6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

Diluent buffer

Incubation time

Incubation temperature:



7) Secondary antibody:

Species:

Reacts against:

Concentration or dilution

Diluent buffer

Incubation time

Incubation temperature:

Fluorochrome or enzyme conjugate:



8) Washing after primary and secondary antibodies:

Buffer

Number of washes



9)Detection method

10) How many times have you run this staining?

Do you obtain the same results every time?

What steps have you altered to try and optimize the use of this antibody?



Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.





Please send this questionnaire by e-mail by cutting and pasting the text in your e-mail (no attachments other than images) to technical@abcam.com.

Read More

Question
Answer

Thanks for your email.

This item is currently back ordered 1-2 weeks from date the order is placed. You should expect to receive this vial of ab64171 in approximately 1 week.

I hope this is helpful. Please contact us again if you have any further questions.

Read More

Answer

Fantastic! Best of luck in your experiments!

Read More

Question
Answer

Thank you for your enquiry.

We have a number of different antibodies against specific acetylated residues on histone H4. However an antibody does not exist that will detect every acetylated residue on histone H4.

You may consider just using one or two antibodies against commonly acetylated histone H4 residues. I'm sure this information is available in the literature.

I hope this is helpful. Please contact me again if you have any further questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (Bone tissue)
Loading amount
25 µg
Specification
Bone tissue
Gel Running Conditions
Reduced Denaturing (15% Gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Dr. Rodrigo Alves

Verified customer

Submitted Apr 17 2012

Question
Answer

DISCOUNT CODE: 100%ABR-T7A9Q
Expiration date: 12th August 2012

I am very pleased to hear you would like to accept our offer and test ab7311 insheep tissue. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview forsheep tissueand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Question
Answer



I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

1-10 of 44 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up