Overview

  • Product nameAnti-acetyl Lysine antibody
    See all acetyl Lysine primary antibodies
  • Description
    Rabbit polyclonal to acetyl Lysine
  • Tested applicationsSuitable for: ELISA, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Synthetic acetylated peptide derived from Lysine-acetlyated proteins (G-A-KA-KA-KA-KA-KA-KA).

  • Positive control
    • Human lung carcinoma tissue and COS7 cell extracts treated with TSA (400uM, 24hours). This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Purification notesab61257 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific acetylated peptide. The antibody against non-acetylated peptide was removed by chromatography using non-acetylated peptide corresponding to the acetylation site.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab61257 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
ELISA 1/10000.
IHC-P 1/50 - 1/100.
WB 1/500 - 1/1000. Detects a band of approximately 85 kDa.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • RelevanceIn 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

Anti-acetyl Lysine antibody images

  • ICC/IF image of ab61257 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab61257 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab61257 at 1/50 - 1/100 dilution staining Acetyl lysine in human lung carcinoma by Immunohistochemistry, Paraffin-embedded tissue, in the absence or presence of the immunising peptide.
  • All lanes : Anti-acetyl Lysine antibody (ab61257) at 1/500 dilution

    Lane 1 : COS7 cell extracts treated with TSA (400uM, 24hours)
    Lane 2 : COS7 cell extracts treated with TSA (400uM, 24hours) with immunising peptide at 5 µg

    Lysates/proteins at 5 µg per lane.


    Observed band size : 25,45,85,120 kDa (why is the actual band size different from the predicted?)

  • Developed using the ECL technique

    Exposure time : 3 minutes

    Image courtesy of an anonymous Abreview.

    293 histone (acid extracted) loaded at 2µg.
    Blocking step performed using 0.3% BSA for 12 hours at 4°C.
    ab61257 used at a 1/1000 dilution for 1 hour at 25°C.
    The secondary used was an HRP conjugated donkey anti-rabbit IgG polyclonal used at a 1/2000 dilution.

    See Abreview

References for Anti-acetyl Lysine antibody (ab61257)

This product has been referenced in:
  • Kuroda N  et al. Apoptotic response through a high mobility box 1 protein-dependent mechanism in LPS/GalN-induced mouse liver failure and glycyrrhizin-mediated inhibition. PLoS One 9:e92884 (2014). IHC-P ; Human . Read more (PubMed: 24690901) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Purified protein (293 histone (acid extracted))
Loading amount 2 µg
Specification 293 histone (acid extracted)
Gel Running Conditions Reduced Denaturing (18 %)
Blocking step BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 0.3% · Temperature: 4°C
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Submitted Dec 12 2011

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