Anti-Histone H1 antibody [1415-1] (ab62884)

Overview

  • Product nameAnti-Histone H1 antibody [1415-1]See all Histone H1 primary antibodies ...
  • Description
    Mouse monoclonal [1415-1] to Histone H1
  • Tested applicationsFlow Cyt, IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Tissue, cells or virus corresponding to Human Histone H1. Immunogen is nuclei of myeloid leukemia biopsy cells.

  • Positive control
    • Human liver nuclear proteins. ICC/IF - HeLA cells

Properties

Applications

Our Abpromise guarantee covers the use of ab62884 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
IF Use a concentration of 2 µg/ml.

For better results, cells should be fixed with methanol.

Flow Cyt Use 0.5µg for 106 cells.
IHC-P Use a concentration of 5 µg/ml.
ICC/IF 1/30.
WB 1/100 - 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 23 kDa).

Target

Anti-Histone H1 antibody [1415-1] images

  • ab62884 staining Histone H1 in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab62884 at 2µg/ml and ab6046 (anti beta Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®084) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • Anti-Histone H1 antibody [1415-1] (ab62884) at 1/100 dilution + human liver nuclear proteins at 30 µg

    Predicted band size : 23 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)
  • IHC image of ab62884 staining in Breast Cancer formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab62884, 5µg/ml, for 15 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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • ab62884 staining Histone H1 in mouse primary hepatocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 3% BSA + 0.1% Triton X-100 for 1 hour at 22°C. Samples were incubated with primary antibody (1/30 in blocking buffer) for 20 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab62884 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab62884, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Histone H1 antibody [1415-1] (ab62884)

This product has been referenced in:
  • Lin CJ  et al. Histone variant H3.3 maintains a decondensed chromatin state essential for mouse preimplantation development. Development 140:3624-34 (2013). ICC/IF ; Mouse . Read more (PubMed: 23903189) »
  • Gastaldello S  et al. Caspase-1 promotes Epstein-Barr virus replication by targeting the large tegument protein deneddylase to the nucleus of productively infected cells. PLoS Pathog 9:e1003664 (2013). WB ; Human . Read more (PubMed: 24130483) »

See all 4 Publications for this product

Product Wall

1. I can confirm that histone H1 antibody ab62884 has been tested with mouse and human samples. However, I am sorry that as far as we are aware, it has never been tested in ChIP. All tested applications and species covered by the 12 month guarantee are...

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Application Western blot
Loading amount 25 µg
Gel Running Conditions Non-reduced Denaturing (15%)
Sample Human Cell lysate - whole cell (Hep3B)
Specification Hep3B
Blocking step I-Block(Applied biosystems) as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
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Submitted Jun 23 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 H3K9ac
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Submitted Mar 03 2014

Application Immunoprecipitation
Immuno-precipitation step Protein G
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Total protein in input 30 µg
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Submitted Feb 27 2014

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
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Submitted Feb 24 2014

I am sorry to confirm that the Anti-Histone H1 antibody [1415-1] - Carboxyterminal end (ab62884) has not been tested in horse. For that reason we can not guarantee it to work.

It would be possible to find a suitable antibody on our website. If...

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Thank you for your reply and for providing this additional information.

Please let me know how the antibody works in the blot that you're running today. We only have full size vials of all of our products so I'm unable to provide a small samp...

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Thank you for your reply and for sending this additional information.

Could you also tell me what dilution of antibody you've tried?

You mentioned that other antibodies worked on the same membrane. Were these antibodies tested with ...

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Thank you for contacting us.

H1.2 specific antibodies,
ab4086; http://www.abcam.com/Histone-H1-2-antibody-ChIP-Grade-ab4086.html. (we however haven't tested the specificity to other forms)
ab17677; http://www.abcam.com/Histone-H1-2-...

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Thank you for contacting us.

ab87556 was used with a sequence from H1FOO_HUMAN protein; http://www.uniprot.org/uniprot/Q8IZA3. According to swissprot the molecular weight is 35 kDa.

ab83058; the sequence used is from P07305 http://...

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