Anti-Histone H1.2 antibody - ChIP Grade (ab4086)

Overview

  • Product nameAnti-Histone H1.2 antibody - ChIP GradeSee all Histone H1.2 primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H1.2 - ChIP Grade
  • SpecificityThis antibody has only been tested on bulk HeLa histones. We expect this antibody to be specific for Histone H1.2 - however we have not tested this specifically on different histone H1 variants.
  • Tested applicationsChIP, WB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide derived from residues 1 - 100 of Human Histone H1.2.

    .

  • Positive control
    • HeLa Histone preparation
  • General notes

    Histone H1.2 is one of five known h1 variants (known as H1.1/2/3/4/5 and/or H1.a/b/c/d/e). The H1 variants differ from each other in the amino acid sequence of their N-terminal regions.

Properties

Applications

Our Abpromise guarantee covers the use of ab4086 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
ChIP Use 2-6 µg for 25 µg of chromatin.
WB 1/500. Detects a band of approximately 25 kDa (predicted molecular weight: 21 kDa).Can be blocked with Human Histone H1.2 peptide (ab16936).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionHistones H1 are necessary for the condensation of nucleosome chains into higher order structures.
  • Sequence similaritiesBelongs to the histone H1/H5 family.
    Contains 1 H15 (linker histone H1/H5 globular) domain.
  • Cellular localizationNucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H1 histone family member 2 antibody
    • H1.a antibody
    • H12_HUMAN antibody
    • H1F2 antibody
    • H1s-1 antibody
    • HIST1H1C antibody
    • Histone 1 H1c antibody
    • Histone cluster 1 H1c antibody
    • Histone H1.2 antibody
    • Histone H1c antibody
    • Histone H1d antibody
    • Histone H1s-1 antibody
    • MGC3992 antibody
    see all

Anti-Histone H1.2 antibody - ChIP Grade images

  • Histone H1.2 - ChIP Grade was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab4086.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 21kDa; Histone H1.2 - ChIP Grade



  • Predicted band size : 21 kDa

    ab4086 Histone H1.2

    The antibody was used at a dilution of 1/500

    Lane 1: HeLa Histone (5ug) + ab4086 
    Lane 2: HeLa Histone (5ug) + ab4086 + 1µg/ml of peptide (Histone H1.2) (ab16936)

    Secondary ab: Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed ab7090 (1/5000)

    Exposure time: 1 minute
    Expected molecular weight: 21.3 kDa

    ab4086 Histone H1.2

    The antibody was used at a dilution of 1/500

    Lane 1: HeLa Histone (5ug) + ab4086
    Lane 2: HeLa Histone (5ug) + ab4086 + 1µg/ml of peptide (Histone H1.2) (ab16936)

    Secondary ab: Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed ab7090 (1/5000)

    Exposure time: 1 minute
    Expected molecular weight: 21.3 kDa
  • ICC/IF image of ab4086 stained human HeLa cells. The cells were PFA fixed (3.7% PFA, 10 min) and incubated with the antibody (ab4086, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • 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  ab4086 (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.
        

  • IHC image of Histone H1.2 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 ab4086, 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.

References for Anti-Histone H1.2 antibody - ChIP Grade (ab4086)

This product has been referenced in:
  • Millán-Ariño L  et al. Mapping of six somatic linker histone H1 variants in human breast cancer cells uncovers specific features of H1.2. Nucleic Acids Res 42:4474-93 (2014). ChIP ; Human . Read more (PubMed: 24476918) »
  • Shan L  et al. GATA3 cooperates with PARP1 to regulate CCND1 transcription through modulating histone H1 incorporation. Oncogene N/A:N/A (2013). Read more (PubMed: 23851505) »

See all 6 Publications for this product

Product Wall

Thank you for contacting us.

Your credit note ID is xxx.

I am sorry that these 2 antibodies did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed a...

Read More

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

Read More

Thank you very much for your telephone enquiry.

I am sorry to confirm that to our knowledge, the two histone H1.2 antibodies we have are regrettably not tested and guaranteed in mouse or frozen sections (IHC-Fr).

ab17677 Anti-Hist...

Read More

Thank you for contacting us. We are pleased to hear that you attended our seminar on the 16th and hope you enjoyed it. I appreciate the time you have spent in the laboratory and understand your concerns. We would like to investigate this particular...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (MRC5VA)
Loading amount 15 µg
Specification MRC5VA
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Sep 01 2009

Thank you for your enquiry. This antibody was tested on purified core histones purchased from another company. They were purified by acid extraction precipitation from log phase untreated HeLa cells. I hope this information helps, please do not ...

Read More

Thank you for your e-mail. We are very pleased to hear that you are satisfied with the antibody ab4086. I enclose our recommended protocol for blocking, please do not hesitate to contact us again if you need further information. In principle, a ...

Read More

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