Anti-acetyl Lysine antibody - ChIP Grade (ab21623)

Overview

  • Product nameAnti-acetyl Lysine antibody - ChIP Grade
    See all acetyl Lysine primary antibodies
  • Description
    Rabbit polyclonal to acetyl Lysine - ChIP Grade
  • SpecificityRecognises proteins acetylated on lysine residues. Tested: acetylated histone, acetylated BSA, and acetylated MBP, no reaction to the non acetylated proteins.
  • Tested applicationsSuitable for: IP, IHC-P, ChIP, ICC/IF, WB, ELISAmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Acetylated KLH conjugates.

  • Positive control
    • Subconfluent MMRU cells with or without HDAC inhibitor treatment
  • General notes


    Use to detect acetylation of lysine.

Properties

Applications

Our Abpromise guarantee covers the use of ab21623 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
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. PubMed: 24160175
ChIP Use 2-4 µg for 25 µg of chromatin.
ICC/IF 1/100 - 1/250. Subconfluent MMRU cells seeded in a 6-well plate with or without HDAC inhibitor treatment were fixed and permeabilized with 1% formaldehyde and 0.5% Triton X-100 for 10 minutes and blocked with 10 mg mL-1 BSA for 1 hour at room temperature. Cells were stained with FITC-conjugated anti-acetylated lysine antibody at 1:100 dilution with PBSt 1 % BSA for 1 hour at room temperature.
WB 1/1000 - 1/2500.
ELISA 1/5000.

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 - ChIP Grade images

  • Primary: All Lanes: Anti acetyl Lysine antibody (ab21623) at 1:1000. Lane 1: Marker. Lane 2: HeLa cells vehicle-treated (ab139414). Lane 3: HeLa cells, trichostatin A-treated (ab139414). Lysates at 20 ug/lane . Secondary: All Lanes: Goat anti-Rabbit IgG 1:10000. Performed under reducing conditions. Blocking buffer: 5% milk in PBS. Observed band sizes: 11 kDa 15kDa 45kDa 50 kDa.
  • Lane 1 = Extract of Mcf7 cells incubated with vehicle ? 20 ug. Lane 2 = Extract of Mcf7 cells incubated with trichostatin A ? 20 ug. Lane 3 = Extract of Mcf7 cells incubated with EX527 ? 20 ug. Lane 4 = Extract of Mcf7 cells incubated with nicotinamide ? 20 ug. Lane 5 = Extract of Mcf7 cells incubated with camptothecin ? 20 ug. Lane 6 = Extract of Mcf7 cells incubated with camptothecin and trichostatin A ? 20 ug. Lane 7 = Extract of Mcf7 cells incubated with camptothecin and EX527 ? 20 ug. Lane 8 = Extract of Mcf7 cells incubated with camptothecin and nicotinamide ? 20 ug. Lane 9 = Extract of 293T cells incubated with vehicle ? 20 ug. Lane 10 = Extract of 293T cells incubated with camptothecin ? 20 ug. Lane 11 = Extract of 293T cells incubated with camptothecin and trichostatin A ? 20 ug. Lane 12 = Extract of 293T cells incubated with camptothecin and EX527 ? 20 ug. Lane 13 = Extract of 293T cells incubated with camptothecin and nicotinamide ? 20 ug
    SDS PAGE performed under reducing conditions (100mM DTT ? Sample heated at 50?C). Primary : Lanes 1-13: Rabbit anti acetyl Lysine antibody (ab21623) at 1/500 dilution. Secondary : Lanes 1-13: Goat anti rabbit IgG(H&L)-IR680 at 1:10,000 (in green). Developed: Oddysey. Blocking: in 5% Milk + PBS for 3 hours at RT. Primary antibody: in 5% BSA + 50mM Tris pH 7.5 + 150 mM NaCl + 0.05% Tween-20. Secondary antibody: in 5% Milk + PBS + 0.1% Tween-20 + 0.01%SDS for 2 hour at RT. Predicted band size : multiple. Observed band size : multiple
  • Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of  ab21623 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.    

  • All lanes : Anti-acetyl Lysine antibody - ChIP Grade (ab21623) at 0.5 µg/ml

    Lane 1 : Untreated Human melanoma cell lysate
    Lane 2 : TSA-treated Human melanoma cell lysate

    Lysates/proteins at 75 µg per lane.

    Secondary
    Goat anti-rabbit IgG HRP at 0.25 µg/ml
    Developed using the ECL technique

    Observed band size : 16-18 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 12-14 kDa. We are unsure as to the identity of these extra bands.
  • Immunofluorescent staining of Human melanoma cells, using Rabbit polyclonal to acetyl Lysine (ab21623) at 1:100 dilution.

    1: Untreated cells

    2: TSA treated cells

  • p53 acetylation upon Doxorubicin treatment in human melanoma cells (MMRU cells). MMRU cells were treated with 0.5 ug/ml Dox for various times and lyzed for whole protein. Immunoprecipitation was performed with ab21623. Western blot was performed to detect the immunoprecipitated p53 with anti-p53 antibody.

References for Anti-acetyl Lysine antibody - ChIP Grade (ab21623)

This product has been referenced in:
  • Lardenois A  et al. The conserved histone deacetylase Rpd3 and its DNA binding subunit Ume6 control dynamic transcript architecture during mitotic growth and meiotic development. Nucleic Acids Res 43:115-28 (2015). Read more (PubMed: 25477386) »
  • Jiang X  et al. Inhibition of HDAC3 promotes ligand-independent PPAR? activation by protein acetylation. J Mol Endocrinol 53:191-200 (2014). WB . Read more (PubMed: 24982244) »

See all 13 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 18 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 18 2016

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12% gel)
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Specification Skeletal muscle
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 09 2014

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Denaturing (12.5)
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 28 2013

Thank you for contacting us.

Technically yes!, the antibody should be able to pull down the peptides with acetylated lysine.

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

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Thank you for contacting us.
The immunogen for ab221623 is chemically acetylated kehole limpet hemocyanin (KLH) and the antibody was affinity purified with Ne-Acetyl-L-lysine on Agarose.
I hope this information is helpful to you. Please do not...

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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...

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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- ...

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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...

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