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ChIP - acetyl Lysine antibody (ab21623)
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Western blot - acetyl Lysine antibody (ab21623)
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Immunofluorescence - acetyl Lysine antibody (ab21623)
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Product Name
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acetyl Lysine antibody - ChIP Grade
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See all acetyl Lysine antibodies (9)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to acetyl Lysine - ChIP Grade
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Immunogen
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Acetylated KLH conjugates.
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Specificity
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Recognises proteins acetylated on lysine residues. Tested: acetylated histone, acetylated BSA, and acetylated MBP, no reaction to the non acetylated proteins.
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Tested applications
(see key)
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ChIP, ELISA, ICC/IF, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ChIP: Use 2-4µg for 25µg of chromatin.
ELISA: 1/5000.
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.
IP: Use at an assay dependent dilution.
WB: 1/1000 - 1/2500.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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Subconfluent MMRU cells with or without HDAC inhibitor treatment
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Research areas
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Chromatin and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Chromatin and Nuclear Signaling >> Histones >> H2A >> Acetylated
Chromatin and Nuclear Signaling >> Histones >> H2B >> Acetylated
Chromatin and Nuclear Signaling >> Histones >> H4 >> Acetylated
Chromatin and Nuclear Signaling >> Histones >> H3 >> Acetylated
Chromatin and Nuclear Signaling >> Chromatin Binding Proteins >> Acetyl Lysine
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Relevance
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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.
Use to detect acetylation of lysine.
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.1% Sodium Azide
Constituents: TBS Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Purification notes
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The antibody was specifically purified with immobilised acetylated lysine on agarose
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Form
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Liquid
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Concentration
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0.250 mg/ml
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Storage instructions
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Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this acetyl Lysine antibody - ChIP Grade is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for acetyl Lysine antibody - ChIP Grade images, references, products related to ab21623 and other tools.
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acetyl Lysine antibody - ChIP Grade images:
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 ChIP - acetyl Lysine antibody (ab21623)
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.
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Western blot - acetyl Lysine antibody (ab21623)
All lanes : 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.
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Immunofluorescence - acetyl Lysine antibody (ab21623)
Immunofluorescent staining of Human melanoma cells, using Rabbit polyclonal to acetyl Lysine (ab21623) at 1:100 dilution. 1: Untreated cells 2: TSA treated cells
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Immunoprecipitation - acetyl Lysine antibody (ab21623)
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.
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References for acetyl Lysine antibody - ChIP Grade (ab21623)
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This product has been used in:
Kim SC et al. Substrate and functional diversity of lysine acetylation revealed by a proteomics survey. Mol Cell 23:607-18 (2006). PubMed: 16916647
If you publish research using ab21623 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to acetyl Lysine
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