Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Anti-5-hydroxymethylcytosine (5-hmC) 抗体 [AB3/63.3] - ChIP Grade (ab106918)

Reviews (2)Q&A (3)References (19)
Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
  • Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
  • MeDIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
  • ChIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)

Key features and details

  • Rat monoclonal [AB3/63.3] to 5-hydroxymethylcytosine (5-hmC) - ChIP Grade
  • Suitable for: ICC/IF, IP, ChIP, IHC-Fr, Dot blot, MeDIP
  • Reacts with: Species independent
  • Isotype: IgG2a

製品の概要

  • 製品名

    Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade
    5-hydroxymethylcytosine (5-hmC) 一次抗体 製品一覧

  • 製品の詳細

    Rat monoclonal [AB3/63.3] to 5-hydroxymethylcytosine (5-hmC) - ChIP Grade

  • 由来種

    Rat

  • アプリケーション

    適用あり: ICC/IF, IP, ChIP, IHC-Fr, Dot blot, MeDIPmore details

  • 種交差性

    交差種: Species independent

  • 免疫原

    5-hydroxymethylcytidine conjugated to KLH

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab106918の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ICC/IF (1)
1/500 - 1/1000.
IP
1/200.
ChIP (1)
Use at an assay dependent concentration.
IHC-Fr
1/500 - 1/1000.
Dot blot
1/500.
MeDIP
Use at an assay dependent concentration.
特記事項
ICC/IF
1/500 - 1/1000.
IP
1/200.
ChIP
Use at an assay dependent concentration.
IHC-Fr
1/500 - 1/1000.
Dot blot
1/500.
MeDIP
Use at an assay dependent concentration.

ターゲット情報

  • 関連性

    5-Hydroxymethylcytosine (5-hmC) is a modified base form of cytosine recently found in Human/mouse brain and inembryonic stem cells. This DNA pyrimidine nitrogen base can be generated by oxidation of 5-methylcytosine, a reaction mediated by the ten-eleven translocation (TET) family of the 5-mC hydroxylases. The function of this base is still not elucidated but it is believed to play an important role in switching genes on and off.

  • 別名

    • 5-hmC antibody

画像

  • Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    Dot blot assay shows that ab106918 specifically recognized 5-hydroxymethyl Cytidine (hmC). Indicated amounts of hmC, methyl Cytidine (mC) and Cytidine (C) were spotted onto a membrane that was then incubated with ab106918. hmC, mC and C were generated in the following way: M13mp18 DNA had been amplified using primers F and R; F: atttccatgagcgtttttcc R: gcaaggcaaagaattagcaa. A 200 uM dNTP end concentration was used with 1. A,G,C,T and 2. A,G,hmC,T; where C had been replaced with HmdCTP. DNA was in vitro methylated with SssI and SAM, and 2ul of pmol of each base was denatured at 95C for 5 min and spotted and dried onto the membrane. The dot blot membrane was blocked with 10%skimmed milk + 1%BSA blocking overnight and then incubated with ab106918 at 1:500 in blocking solution. A goat anti rat HRP secondary antibody was used for ECL detection. This image is from an anonymous collaborator.
  • Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    Dot blot competition assay in which ab106918 was preincubated with 5-hydroxymethyl Cytidine (5hmC) at amounts indicated in figure. Specified amounts of 5hmC, methyl Cytidine (mC) and Cytidine (C) were spotted onto membranes and were then incubated with ab106918 that had been preincubated with 5hmC as shown in figure. ab106918 specifically recognized 5hmC and this was blocked by preincubation with 5hmC at 5 pmol/ug ab106918 (Ab). This image is from an anonymous collaborator.
  • MeDIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    MeDIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    The specificity of ab106918 was confirmed by (h)MeDIP using qPCR validation of regions in ES cells that are highly enriched in 5-hydroxymethyl Cytidine (5hmC) (Pic1, Pic3 and Pic7) or not (Pic5 and Pic10). This image is from an anonymous collaborator.
  • ChIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)
    ChIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (ab106918)This image is courtesy of an anonymous Abreview
    ChIP analysis of mouse ES nuclear cell lysate using ab106918 to bind 5-hydroxymethyl Cytidine. Chromatin was obtained by incubating with primary antibody (0.5 µg/µg chromatin in a glycerol IP buffer) for 16 hours at 4°C. Protein binding was detected using real-time PCR.

    See Abreview

参考文献 (19)

ab106918 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab106918 は 19 報の論文で使用されています。

  • Feng W  et al. DHX33 Recruits Gadd45a To Cause DNA Demethylation and Regulates a Subset of Gene Transcription. Mol Cell Biol 40:N/A (2020). PubMed: 32312884
  • Zhang X  et al. TET2 suppresses nasopharyngeal carcinoma progression by inhibiting glycolysis metabolism. Cancer Cell Int 20:363 (2020). PubMed: 32774157
  • Park H  et al. Rad50 mediates DNA demethylation to establish pluripotent reprogramming. Exp Mol Med 52:1116-1127 (2020). PubMed: 32665583
  • Cakouros D  et al. Specific functions of TET1 and TET2 in regulating mesenchymal cell lineage determination. Epigenetics Chromatin 12:3 (2019). PubMed: 30606231
  • Wang J  et al. Characterization of DNA hydroxymethylation profile in cervical cancer. Artif Cells Nanomed Biotechnol 47:2706-2714 (2019). PubMed: 31271297
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-5 of 5 Abreviews or Q&A

ChIP abreview for Anti-5-hydroxymethyl Cytidine antibody [AB3/63.3]

 Excellent
Abreviews
abreview image
Application
ChIP
Sample
Mouse Cell lysate - nuclear (Mouse ES cells)
Specification
Mouse ES cells
Type
Native ChIP (N-ChIP)
Detection step
Real-time PCR
Positive control
Taf5
Negative control
Magoh2
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Dec 06 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-5-hydroxymethyl Cytidine antibody [AB3/63.3]

 Poor
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Kirk Mcmanus

Verified customer

投稿 Feb 10 2012

Question

I am interesting to analyze the methylation and hydroximethylation status of my samples (adherent cells) and i found our products: ab117133 and ab117135.

Are these kits suitable for RT-PCR analysis?Do the 24 or 48 tests correspond to 24 or 48 single immunoprecipitations?For example, if i have to analyze two different cell type, i have to consume 6 tests (two for 5mc, two for negative control IgG and two for imput DNA)?And finally, I am interesting to hydroximethylation, so do you have similar kit for this modification?If i purchase the anti-5-hydroxymethyl Cytidine antibody (ab106918), i could use this antibody and so different negative control (in place of Anti-5-methylcytosine and mouse IgG) with the same kit?

Read More
Answer

Both of these kits (ab117133 and ab117135) will immunoprecipitate methylated DNA. The ab117133 Kit uses purified DNA as starting materials while the ab117135 Kit uses cells as starting materials. I can confirm that the immunoprecipitated products are suitable to use for RT-PCR.

The 24 or 48 tests correspond to single immunoprecipitations and not to the number of samples that you are able to assay. Therefore, if for one cell type you need to do 6 tests, then you will be able to run 4 cell types with a 24 sample kit. The kits are in strip format though, so you can save what you don't use for a later time.

With regards to a hydroxymethylated DNA kit, we have https://www.abcam.com/episeeker-hydroxymethylated-dna-quantification-kit-colorimetric-ab117130.html, Seeker hydroxymethylated DNA Quantification Kit (Fluorometric) (ab117131) or https://www.abcam.com/episeeker-hydroxymethylated-dna-immunoprecipitation-hmedip-kit-ab117134.html.

The more suitable kit would depend on what type of information you want from your samples. The ab117130 and ab117131 kits will allow you to look at global hydroxymethylation levels of their samples and would tell you the percent of hydroxymethylation of your DNA but would not allow for RT-PCR. The kit ab117134 will allow the you to explore regions of hydroxymethylated DNA by immunoprecipitating/enriching the hydroxymethylated DNA followed by RT-PCR or hMeDIP-chip which is maybe more useful to you.

You can indeed purchase the anti-5-hydroxymethyl cytidine antibody (ab106918) and use it with the 5-mC immunoprecipitation kit since you coat the antibody on the wells.

Read More

Question

Dear Tech support

One of our customers wishes to know if antibodies ab10805, ab106918 are suitable ad guaranteed for Vibritome processed and PFA fixated tissue samples?

Please advise.

Thank you for your kind help.

Regards,

Read More
Answer

Thank you for contacting us.


Anti-5-Methyl Cytidine antibody (ab10805) has been tested and is guaranteed in paraffin embedded and frozen section immunohistochemistry. Anti-5-hydroxymethyl Cytidine antibody (ab106918) has only been tested in frozen sections and would be guaranteed for this.


We have no information on if they will work with Vibritome processed sections. If your customer would like, I can issue a testing discount for ab106918 to be used in immunohistochemical staining of paraffin embedded tissue sections. In order to process this I will need their email address and the institution they belong to.


I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

Read More

Question

Dear Tech support

One of our customers in interested in ab106918.

The antibody is not written to have been tested for IHC-P, would you like the customer to order it as part of the abreview promotion?

Otherwise, could it be guaranteed to work for IHC-P and is there a protocol available?

Thank you for your kind help.

Regards,

Read More
Answer

Thank you for contacting us.

To our knowledge, ab106918 has not yet been tested in IHC-P. Your customer would therefore qualify for the testing discount promotion. If your customer would like to take part in this scheme please could you provide the following details and I will issue the code:

End user email address:
End user institution:

The Terms and Conditions of this offer can be found at:


https://www.abcam.com/collaborationdiscount


If you have any further questions please do not hesitateto contact us again.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com