Name: Anti-Histone H1.0 antibody [EPR6537]
Brand: RabMAb
SKU: ab125027

Product name

Anti-Histone H1.0 antibody [EPR6537]
See all Histone H1.0 primary antibodies
Description

Rabbit monoclonal [EPR6537] to Histone H1.0
Host species

Rabbit
Tested applications

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

Reacts with: Human
Immunogen

Synthetic peptide within Human Histone H1.0 aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P07305
Positive control
- WB: BxPC3 and fetal Human lung lysates IHC-P: Human kidney tissue ICC/IF: A431 cells
General notes

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR6537
Isotype

IgG
Research areas

Alternative Versions
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Histone H1.0 protein (ab158626)

Our Abpromise guarantee covers the use of ab125027 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
WB		1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 21 kDa).
IHC-P		1/100 - 1/400. Antigen retrieval is not essential but may optimise staining.
ICC/IF		1/100 - 1/250.

Function

Histones H1 are necessary for the condensation of nucleosome chains into higher-order structures. The H1F0 histones are found in cells that are in terminal stages of differentiation or that have low rates of cell division.
Sequence similarities

Belongs to the histone H1/H5 family.
Contains 1 H15 (linker histone H1/H5 globular) domain.
Post-translational
modifications

Phosphorylated on Ser-17 in RNA edited version.
Cellular localization

Nucleus. Chromosome. The RNA edited version has been localized to nuclear speckles. During mitosis, it appears in the vicinity of condensed chromosomes.
Target information above from: UniProt accession P07305 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3005 Human
- Omim: 142708 Human
- SwissProt: P07305 Human
- Unigene: 745024 Human
Alternative names
- H1 histone family member 0 antibody
- H1(0) antibody
- H10 antibody
- H10_HUMAN antibody
- h1f0 antibody
- H1FV antibody
- Histone H1'' antibody
- Histone H1(0) antibody
- Histone H1.0 antibody
- Histone H10 antibody
- Histone H5 antibody
- MGC5241 antibody
- N-terminally processed antibody
see all

Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

All lanes : Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 10 µg (purified)

Lane 1 : human fetal lung lysate
Lane 2 : BxPC-3 lysate
Lane 3 : A431 lysate
Lane 4 : human fetal kidney lysate

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 21 kDa
Observed band size: 32 kDa
why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Immunofluorescence staining of BxPC-3 cells with purified ab125027 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab125027 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Immunohistochemical staining of paraffin embedded rat kidney with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Immunohistochemical staining of paraffin embedded mouse liver with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Immunohistochemical staining of paraffin embedded human thyroid carcinoma with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Immunohistochemical staining of paraffin embedded human pancreas with purified ab125027 at a working dilution of 1/400. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

All lanes : Anti-Histone H1.0 antibody [EPR6537] (ab125027) at 1/1000 dilution (unpurified)

Lane 1 : BxPC3 lysates
Lane 2 : Human fetal lung lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti rabbit at 1/2000 dilution

Predicted band size: 21 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Unpurified ab125027, at a 1/100 dilution, staining Histone H1 in paraffin embedded Human kidney tissue by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.0 antibody [EPR6537] (ab125027)

Unpurified ab125027, at a 1/100 dilution, staining Histone H1 in A431 cells by Immunofluorescence.

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

Datasheet

This product has been referenced in:

Chakrabarti J et al. Hedgehog signaling induces PD-L1 expression and tumor cell proliferation in gastric cancer. Oncotarget 9:37439-37457 (2018). Read more (PubMed: 30647844) »
Jiang P et al. Histones facilitate a-synuclein aggregation during neuronal apoptosis. Acta Neuropathol 133:547-558 (2017). Read more (PubMed: 28004278) »

See all 7 Publications for this product

Publishing research using ab125027? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

Thank you for your response. Histone H1.2, 1.3 or 1.4 are about 21-22KD. Which of your antibody recognize these three isoforms, especially H1.4?
I also notice that you have Anti-Histone H1 (tri methyl K25) antibody (ab17347) which detects H1.4K25. But there is a post on your web site (question 2) states:"This antibody may not recognize methylated H1 but only the H3." Do you have any update for it? I am
interested to purchase this antibody if it indeed can recognize H1.4K25(K26).

Abcam community

Verified customer

Asked on Mar 06 2012

Answer

Thank you for contacting us.

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

H1.3 abs
ab24174; https://www.abcam.com/Histone-H1-3-antibody-ab24174.html
H1.4
ab105522; https://www.abcam.com/Histone-H1-4-antibody-ab105522.html
ab116393; https://www.abcam.com/Histone-H1-4-antibody-ab116393.html

The antibody ab17347 is for tri methylated H1, unfortunately it also cross react with methylated histone H3 and other isoform of H1. This antibody however will not recognize di and mono methylated histone H1. Is this the answer to your question? Let me know if you any further question about the specificity of these products.

Abcam Scientific Support

Answered on Mar 06 2012

Question

Would you please help to verify the following questions regarding your
anti-histone H1 anti bodies:
Anti-Histone H1 antibody (ab87556): Detects a ˜32KD band;
Anti-Histone H1 antibody (ab83058) : Detects a ˜24KD band;
Anti-Histone H1 antibody [1415-1] - Carboxyterminal end (ab62884) :
Detects a ˜17KD band;
Anti-Histone H1 antibody [AE-4] (ab71594) : no WB image;
Anti-Histone H1 antibody [EPR6537] (ab125027): WB ˜32KD
1). Do they represent different modifications of H1 or different isofroms
of H1, since the band sizes recognized are so diffenrent? (But MW of H1
isoforms are around 22KD).
2). Will above antibodies also cross-react with other histones like H3 or H4?
Anti-Histone H1 antibody [0.T.60] (ab4269): Detects isoforms 0-5,
˜25-32KD; Why the sizes are different from the oredicted sizes, or due to
modification or something else?
In the end, we need an antibody to detect Histone H1 in human cell lines
such as HEK293 and SH-SY5Y, which one do you recommend?
Thank you!

Abcam community

Verified customer

Asked on Mar 05 2012

Answer

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://www.uniprot.org/uniprot/P07305. The molecular weight is ˜ 21kDa.

ab71594 was used with a sequence from H1FOO_HUMAN protein; http://www.uniprot.org/uniprot/Q8IZA3. According to swissprot the molecular weight is ˜ 35 kDa.

ab125027 was used with a sequence from H1FOO_HUMAN protein; http://www.uniprot.org/uniprot/Q8IZA3. According to swissprot the molecular weight is ˜ 35 kDa.

These antibodies are specific to human H1 protein only not the modifications. You can purchase any of these to detect H1 human protein.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Mar 05 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Recombinant Anti-Histone H1.0 antibody [EPR6537] (ab125027)

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References

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