Overview

  • Product name
    Anti-Methylated Lysine (di methyl , mono methyl ) antibody
    See all Methylated Lysine primary antibodies
  • Description
    Rabbit polyclonal to Methylated Lysine (di methyl , mono methyl )
  • Host species
    Rabbit
  • Specificity
    Recognize proteins methylated on lysine residues (mono- & di-methyl lysine). This antibody was affinity purified with mono-methyl lysine and di-methyl lysine, and may also react with tri-methyl lysine. Does not cross-react with acetylated protein.
  • Tested applications
    Suitable for: IHC-P, WB, ELISAmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Methylated Lysine conjugated to KLH.

  • Positive control
    • HCL (0.1N) extracted and acetone precipited histone fraction

Properties

Applications

Our Abpromise guarantee covers the use of ab23366 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
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 1 µg/ml. Do not use milk for any of the buffers as complex milk proteins may contain competitive methylated proteins. Use BSA instead. ab23366 incubation time 2h RT. If no or weak signal is obtained overnight at 4deg might help.
ELISA Use at an assay dependent concentration.

Target

  • Relevance
    Many proteins are post translationally modified. Modifications such as phosphorylation, glycosylation, ubiquitination and methylation have been shown to play an important role in the development, physiology and disease of animals and plants. Histone lysine methylation is regarded as a very stable modification with important functions in epigenetic gene control and for organizing chromatin domains.
  • Alternative names
    • Dimethylysine antibody
    • Methyl lysine antibody
    • N epsilon dimethyl lysine antibody
    • N epsilon mono methyllysine antibody
    • N epsilon trimethyl lysine antibody
    • N(epsilon) trimethyl lysine antibody
    • Trimethyl lysine antibody
    see all

Images

  • Lane 1 : Anti-Methylated Lysine (tri methyl) antibody (does not cross react with mono, or di methyl lysine residues)
    Lane 2 : Anti-Methylated Lysine (di methyl , mono methyl ) antibody (ab23366)

    All lanes : Yeast extract
  • ab23366 at 1/200 staining human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed a heat mediated antigen retrieval step was performed in citrate buffer. An HRP conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • ab23366 staining formalin fixed paraffin-embedded rat kidney tissue sections.  The section was subjected to heat mediated antigen retrieval in citrate buffer (pH 6.0) and blocked with 5% serum for 30 minutes at 20°C, prior to incubation with the primary antibody (diluted 1/200 in PBS + 5% serum) for 45 mintes at 20°C.  An HRP-conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Lee SW  et al. MicroRNAs Overcome Cell Fate Barrier by Reducing EZH2-Controlled REST Stability during Neuronal Conversion of Human Adult Fibroblasts. Dev Cell 46:73-84.e7 (2018). Read more (PubMed: 29974865) »
  • Feldman M & Levy D Peptide inhibition of the SETD6 methyltransferase catalytic activity. Oncotarget 9:4875-4885 (2018). Read more (PubMed: 29435148) »
See all 24 Publications for this product

Customer reviews and Q&As

1-10 of 24 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Hek293 - kidney cell line)
Total protein in input
1000 µg
Immuno-precipitation step
Other - Dynabeads or Protein G-coated magnetic beads
Specification
Hek293 - kidney cell line

Byron Baron

Verified customer

Submitted Sep 06 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Hep3B)
Gel Running Conditions
Non-reduced Denaturing (15%)
Loading amount
20 µg
Specification
Hep3B
Blocking step
I-Block(Applied biosystems) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 03 2017

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Human Cell lysate - whole cell (Human Embryonic Kidney 293 cells)
Specification
Human Embryonic Kidney 293 cells
Total protein in input
1000 µg

Dr. Yi-Wen Chang

Verified customer

Submitted Jan 26 2015

Answer


The short answer is that there is no predicted MW for this antibody to detect in WB. This antibody will detect any protein of any size that has a methylated lysine. Therefore, depending on your sample you may see no bands, many bands, a smear, etc.

Take a look at this reference. I don't have access to the journal so I can't see if there is a WB image, but we know this ab was used in WB here: http://www.ncbi.nlm.nih.gov/pubmed/21204252?dopt=Abstract

Read More

Answer

Thank you for contacting us.

These antibodies are raised against targets that are not species specific. The antibodies would detect the proteins irrespective of species so the Abtrial discount will not be valid.

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

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Answer

Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More

Answer

Thank you for your reply.

I can confirm that ab23366 does not recognize free BSA or BSA-linked with methylarginine.

I hope this information is helpful and wish you good luck with your experiments.

Read More

Answer

Thank you for your inquiry.

I apologize for the delay in my answer. It took some time to pull the data for lot GR32966-7.

I am happy to confirm that we tested lot GR32966-7 not in a peptide array buton the single amino acid linked to BSA such as mono-mehtyllysine-BSA, di-methyllysine-BSA, tru-methyllysine-BSA.

The ELISA titer (OD50) is 80 ng/mL, 200 ng/mL, 50 ng/mL for mono-methyllysine, di-methyllysine and trimethyllysine-BSA, respectively for lot GR32966-7.

We do not know what amino acid affect the binding, presumably the amino acid with a bulky docking side chain such as tyrosine, phenylalanine, isoleucine may contribute to the steric hindrance of the binding.

I hope this information is helpful and wish you good luck with your research.

Read More

Answer

Thank you for your reply.

Based on the available information, any of those antibodies would work for you, but I would recommend ab21623.

If there is anything else I can help you with, please let me know.

Read More

Answer

Thank you for contacting Abcam.

Please find the answers to your question below:

1 - For ab21623, it will recognize proteins acetylated on lysine residues. For ab23366, it will recognize proteins methylated on lysine residues (mono- & di-methyl lysine).

2 - The mutations should not affect the antibodies ability to recognize the Lysine sites.

3 - The immunogen sequence of these antibodies is proprietary information and so I am unable to supply you with those sequences.

If there is anything else I can help you with, please let me know.

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1-10 of 24 Abreviews or Q&A

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