Anti-Methylated Lysine (di methyl) antibody (ab23366)

Overview

  • Product name
    Anti-Methylated Lysine (di methyl) antibody
    See all Methylated Lysine primary antibodies
  • Description
    Rabbit polyclonal to Methylated Lysine (di methyl)
  • 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) antibody (ab23366)

    Lane 1 : Yeast extract
    Lane 2 : 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:
  • Zhang Y  et al. The transcription factor GATA1 and the histone methyltransferase SET7 interact to promote VEGF-mediated angiogenesis and tumor growth and predict clinical outcome of breast cancer. Oncotarget 7:9859-75 (2016). Read more (PubMed: 26848522) »
  • Kaur I  et al. Widespread occurrence of lysine methylation in Plasmodium falciparum proteins at asexual blood stages. Sci Rep 6:35432 (2016). WB, IP, ICC/IF . Read more (PubMed: 27762281) »

See all 17 Publications for this product

Customer reviews and Q&As

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
Username

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
Username

Dr. Yi-Wen Chang

Verified customer

Submitted Jan 26 2015


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, e...

Read More

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 informatio...

Read More

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 guaran...

Read More

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.

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 t...

Read More

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.

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- ...

Read More

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

Read More

1-10 of 23 Abreviews or Q&A

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

Sign up