Overview

  • Product name
    Anti-acetyl Lysine antibody
    See all acetyl Lysine primary antibodies
  • Description
    Rabbit polyclonal to acetyl Lysine
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ELISA, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Acetylated KLH Conjugates

  • Positive control
    • TSA treated mouse spleen cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituent: Tris buffered saline
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab80178 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 Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-P 1/100. (see Abreview)

Target

  • Relevance
    In the nucleus, DNA is tightly packed into nucleosomes generating an environment which is highly repressive towards DNA processes such as transcription. Acetylation of lysine residues within proteins has emerged as an important mechanism used by cells to overcome this repression. The acetylation of non-histone proteins such as transcription factors, as well as histones appears to be involved in this process. Acetylation may result in structural transitions as well as specific signaling within discrete chromatin domains. The role of acetylation in intracellular signaling has been inferred from the binding of acetylated peptides by the conserved bromodomain. Furthermore, recent findings suggest that bromodomain/acetylated-lysine recognition can serve as a regulatory mechanism in protein-protein interactions in numerous cellular processes such as chromatin remodeling and transcriptional activation. The reversible lysine acetylation of histones and non-histone proteins plays a vital role in the regulation of many cellular processes including chromatin dynamics and transcription, gene silencing, cell cycle progression, apoptosis, differentiation, DNA replication, DNA repair, nuclear import, and neuronal repression. More than 20 acetyltransferases and 18 deacetylases have been identified so far, but the mechanistic details of substrate selection and site specificity of these enzymes remain unclear. Over 40 transcription factors and 30 other nuclear, cytoplasmic, bacterial, and viral proteins have been shown to be acetylated in vivo.
  • Alternative names
    • pan acetyl Lysine antibody

Images

  • Immunocytochemistry/ Immunofluorescence analysis of Heat Shocked HeLa Cells labeling acetyl Lysine with ab80178 at 1/100 dilution. Cells were fixed with 2% Formaldehyde for 20 min at RT. DAPI (blue) nuclear counter stain at 1/40000 for 2 hours at RT. A FITC conjugated Goat Anti-Rabbit secondary antibody (green) was used at 1/200 dilution. Localization: Nucleus and Cytoplasm.

    (A) DAPI (blue) nuclear stain. (B) Anti-acetyl Lysine antibody (ab80178) (C) Composite. 

  • ab80178 staining acetyl Lysine in human HepG2 cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol and then blocked using 2% serum for 30 minutes at 25°C. Samples were then incubated with primary antibody at 1/150 for 1 hour at 25°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 594 (red) used at a 1/1000 dilution.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of Heat Shocked HeLa Cells labeling acetyl Lysine with ab80178 at 1/100 dilution. Cells were fixed with 2% Formaldehyde for 20 min at RT. DAPI (blue) nuclear counter stain at 1/40000 for 2 hours at RT. A R-PE conjugated Goat Anti-Rabbit secondary antibody (yellow) was used at 1/200 dilution. Localization: Nucleus and Cytoplasm.

    (A) DAPI (blue) nuclear stain. (B) Anti-acetyl Lysine antibody (ab80178) (C) Composite. 

  • Anti-acetyl Lysine antibody (ab80178) + Cell lysates prepared from TSA treated mouse spleen cells


    Western blot analysis of Mouse Spleen lysates showing detection of Acetylated Lysine protein using Primary Antibody: Rabbit Anti-Acetyl Lysine Polyclonal Antibody (ab80178) at 1:1000.

  • ab80178 staining acetyl Lysine in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval in citric buffer, pH 6.0 prior to blocking with 10% serum for 1 hour at 20°C. The primary antibody was diluted 1/100 and incubated with the sample for 12 hour at 4°C. An HRP-conjugated goat anti-rabbit polyclonal was used as the secondary antibody, diluted 1/200.

    See Abreview

  • ab80178 staining acetyl Lysine (green) in the neuromuscular junction of fruit fly cells by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in formaldehde, permeabilized using 0.4% Triton-X, blocked with 10% NGS for 1 hour at 20°C, then incubated with ab80178 at a 1/200 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal, used at a 1/1000 dilution.

    See Abreview

References

This product has been referenced in:
  • Subbaramaiah K  et al. Prostaglandin E2 down-regulates sirtuin 1 (SIRT1), leading to elevated levels of aromatase, providing insights into the obesity-breast cancer connection. J Biol Chem 294:361-371 (2019). Read more (PubMed: 30409902) »
  • Wang S  et al. Niacin Inhibits Apoptosis and Rescues Premature Ovarian Failure. Cell Physiol Biochem 50:2060-2070 (2018). Read more (PubMed: 30415247) »
See all 17 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Mar 20 2018

Application
Western blot
Sample
Human Purified protein (Vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
100 uM H2O2 for 2 hours
Specification
Vascular smooth muscle cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted May 22 2017

Answer

Thank you for your reply.

Since the antibody you bought is not from our catalogue, I cannot comment of the suitability for your planned experiments.

If this antibody is tested and guaranteed (as we do with our antibodies) for IP and CHIP, then this antibody should most probablybe fine. But of course I will not be able to guarantee this and therefore suggest to get in contact with the company that produces this antibody and requestmore information on it.

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

Read More

Answer

Thank you for your inquiry.

I am assuming that before subjecting the samples to LC-MS/MS, the enrichments of the acetylated lysine containing peptide is planned to be done with immunoprecipitation?

I am happy to confirm that we offer two antibodies specific for acetylated lysine that are tested and guaranteed for IP (ab21623 is also CHIP tested and guaranteed).

Please follow these links to review the respective datasheets:

ab21623

https://www.abcam.com/index.html?datasheet=21623 (or use the following: https://www.abcam.com/index.html?datasheet=21623).

ab80178

https://www.abcam.com/index.html?datasheet=80178 (or use the following: https://www.abcam.com/index.html?datasheet=80178).

We also have extensive protocols for IP on our homepage:

https://www.abcam.com/index.html?pageconfig=resource&rid=11385

https://www.abcam.com/index.html?pageconfig=resource&rid=11382

I am not especially familiar with the requirements that samples have to adhere to for LC/MS/MS. But maybe elution of protein with little antibody contamination is preferable? Therefore I suggest the following protocol:

https://www.abcam.com/index.html?pageconfig=resource&rid=11415

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

Read More

Answer

Thank you for contacting us. We do not currently offer an Anti-acetyl Lysine antibody pre-bound to beads for IP. However, our product ab80178 Anti-acetyl Lysine antibody, a rabbit polyclonal that has been tested in IP. This could be easily bound to Protein A or to Protein G beads. The antibody is covered under our Abpromise for six months and is guaranteed to work in IP. If you should encounter any difficulties when using this product, if we cannot resolve the issue you are having with the antibody then we would be happy to either send a replacement antibody or to process a refund. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your phone call yesterday and for your patience while I have been in touch with the lab regarding this antibody in IP. Unfortunately we do not have any in-house IP data, and I've looked through the references however I couldn't find much information about the protocol. My contact at the lab suggested starting with 5-10uL antibody bound to beads and incubating overnight. If you expect very little acetylated protein in your samples, you may need to use more than this, however. I am sorry that I don't have more specific information for you, but if there is anything else that we can do for you please let me know and I'll be happy to help.    

Read More

Answer

Thank you for your call today and for your questions about ab80178. I have forwarded your enquiry to the testing lab to see if they have any additional information. I will get back to you as soon as I hear from the lab. Please let me know if you have any further questions or if there is anything else that we can do for you.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Fruit fly (Drosophila melanogaster) Cell (Neuromuscular junction)
Specification
Neuromuscular junction
Fixative
Formaldehyde
Permeabilization
Yes - 0,4% Triton-X
Blocking step
0,25% BSA + 10% NGS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 11 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Liver)
Specification
Liver
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer, pH6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 28 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human hepatoma HepG2)
Specification
human hepatoma HepG2
Fixative
Methanol
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C

Dr. Natasha Snider

Verified customer

Submitted Mar 16 2011

1-10 of 11 Abreviews or Q&A

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