Overview

  • Product name

    Anti-Histone H1.2 antibody [EPR12690]
    See all Histone H1.2 primary antibodies
  • Description

    Rabbit monoclonal [EPR12690] to Histone H1.2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Histone H1.2 aa 1-100. The exact sequence is proprietary.
    Database link: P16403

  • Positive control

    • HeLa , A375, 293 and MCF7 cell lysates; Human ovarian carcinoma tissue; Mouse muscle tissue; MCF7 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab181973 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 at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 21 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.

Target

Images

  • All lanes : Anti-Histone H1.2 antibody [EPR12690] (ab181973) at 1/10000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : A375 cell lysate
    Lane 3 : 293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 21 kDa
    Observed band size: 29 kDa
    why is the actual band size different from the predicted?

  • ChIP analysis using ab181973 binding Histone H1.2 in mouse primary hepatocytes. Cells were cross-linked for 10 minutes with 1% formaldehyde. Samples were incubated with the primary antibody (1/100) for 20 hours at 4°C. Protein binding was detected using real-time PCR.
    Negative Control: IgG.

    See Abreview

  • ab181973 staining Histone H1.2 in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

    Control: PBS only

  • Anti-Histone H1.2 antibody [EPR12690] (ab181973) at 1/2000 dilution + MCF7 cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 21 kDa
    Observed band size: 29 kDa why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling Histone H1.2 with ab181973 at 1/100 dilution, followed by prediluted ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse muscle tissue labeling Histone H1.2 with ab181973 at 1/100 dilution, followed by prediluted ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed MCF7 cells labeling Histone H1.2 with ab181973 at 1/250 dilution, followed by Goat anti rabbit IgG (Dylight 488) at 1/200 dilution (green), Dapi staining (blue).

References

This product has been referenced in:

  • Hangas A  et al. Ciprofloxacin impairs mitochondrial DNA replication initiation through inhibition of Topoisomerase 2. Nucleic Acids Res 46:9625-9636 (2018). Read more (PubMed: 30169847) »
See 1 Publication for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (fibroblasts/embryos)
Permeabilization
Yes - Triton 100X - 0.5%
Specification
fibroblasts/embryos
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Dr. Vilceu Bordignon

Verified customer

Submitted Dec 14 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
3%BSA+0.1% Triton X100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
Sample
Mouse Cell (primary hepatoctyes)
Specification
primary hepatoctyes
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 26 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing
Sample
Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification
primary hepatoctyes
Blocking step
I-Block(Applied biosystems) as blocking agent for 30 minute(s) · Concentration: 2µg/mL · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 11 2014

Application
Immunoprecipitation
Immuno-precipitation step
Protein A
Sample
Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification
primary hepatoctyes
Total protein in input
25 µg

Abcam user community

Verified customer

Submitted Mar 05 2014

Application
ChIP
Detection step
Real-time PCR
Sample
Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification
primary hepatoctyes
Negative control
IgG
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
Positive control
no

Abcam user community

Verified customer

Submitted Mar 03 2014

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