Overview

  • Product name
    Anti-Histone H1.2 antibody
    See all Histone H1.2 primary antibodies
  • Description
    Rabbit polyclonal to Histone H1.2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1.2.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • This antibody gave a positive signal in Human H1.2 Recombinant Protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab17677 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
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/1200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use a concentration of 5 µg/ml.
WB 1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).

Target

  • Function
    Histones H1 are necessary for the condensation of nucleosome chains into higher order structures.
  • Sequence similarities
    Belongs to the histone H1/H5 family.
    Contains 1 H15 (linker histone H1/H5 globular) domain.
  • Cellular localization
    Nucleus. 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

Images

  • Histone H1.2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H1.2 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 ab17677.

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

    Band: 35kDa; Histone H1.2

  • All lanes : Anti-Histone H1.2 antibody (ab17677) at 1/1000 dilution

    Lane 1 : Recombinant histone H1
    Lane 2 : Recombinant histone H1.1
    Lane 3 : Recombinant histone H1.2
    Lane 4 : Recombinant histone H1.3
    Lane 5 : Recombinant histone H1.4
    Lane 6 : Recombinant histone H1.5

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

  • Image courtesy of Human Protein Atlas

    ab17677 staining histone H1.2 in human testis, showing a distinct and strong staining pattern in ductus seminiferus and leydig cells. Paraffin embedded human skin tissue was incubated with ab17677 (1/1200 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab17677 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    .
  • ICC/IF image of ab17677 stained human MCF7 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab17677, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and HepG2 cells.

  • All lanes : Anti-Histone H1.2 antibody (ab17677) at 1 µg/ml

    Lane 1 : Histone H1 Recombinant Protein
    Lane 2 : Histone H1.2 (Human) - Recombinant Protein
    Lane 3 : Histone H1 Recombinant Protein with Human Histone H1.2 peptide (ab18500) at 1 µg/ml
    Lane 4 : Histone H1.2 (Human) - Recombinant Protein with Human Histone H1.2 peptide (ab18500) at 1 µg/ml

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 12 minutes

References

This product has been referenced in:
  • Bernstein DI  et al. Genetic variants with gene regulatory effects are associated with diisocyanate-induced asthma. J Allergy Clin Immunol 142:959-969 (2018). Read more (PubMed: 29969634) »
  • Li Z  et al. Destabilization of linker histone H1.2 is essential for ATM activation and DNA damage repair. Cell Res 28:756-770 (2018). Read more (PubMed: 29844578) »
See all 23 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for contacting us and sharing the results you have observed with the anti-Histone H1.2 antibody (ab17677).
Although the Histone H1.2 is expected to be observed at ˜29 kDa, this size can vary depending on the post-translational modifications and the conditions under which the protein gel was performed. Protein migration within a gel can vary quite widely depending on the gel used, the loading buffer and the electrophoresis buffers. The band you have detected at just above 35 kDa is therefore consistent with what could be expected. This is also supported by the very clean blot produced with only one band and with no discernible non-specific binding of the antibody. We have ourselves observed bands of 29-32 kDa in size.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549 lung carcinoma)
Specification
A549 lung carcinoma
Fixative
Formaldehyde
Permeabilization
Yes - 1% TX-100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 24 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (Human B chronic lymphocytic leukaemia cells)
Loading amount
18 µg
Specification
Human B chronic lymphocytic leukaemia cells
Gel Running Conditions
Reduced Denaturing (12 % Tris Glycine Gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Feb 14 2008

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

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