Anti-acetyl Lysine抗体[1C6] - ChIP Grade (ab22550)

概述

  • 产品名称Anti-acetyl Lysine抗体[1C6] - ChIP Grade
    参阅全部 acetyl Lysine 一抗
  • 描述
    小鼠单克隆抗体[1C6] to acetyl Lysine - ChIP Grade
  • 特异性ab22550 recognises proteins with acetylated lysine.
  • 经测试应用适用于: IP, ChIP, ICC/IF, WBmore details
  • 种属反应性
    与反应: Species independent
  • 免疫原

    Synthetic peptide: sequence surrounding the acetylated lysine 9 of histone H3

  • 阳性对照
    • HeLa cell lysate. MCF7 cell line.

性能

应用

Our Abpromise guarantee covers the use of ab22550 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IP Use a concentration of 3 µg/ml.
ChIP Use a concentration of 10 µg/ml.
ICC/IF 1/100 - 1/500.
WB 1/500 - 1/2000.

靶标

  • 相关性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.
  • 别名
    • pan acetyl Lysine antibody

Anti-acetyl Lysine antibody [1C6] - ChIP Grade 图像

  • Immunoflouroescence analysis of HeLa Cells labelling lysine acetylated proteins with ab22550. Formalin fixed cells were permeabalized with 0.1& Triton X-100 in TBS for 10 mins at room temperature and subsequently blocked with BSA at room temperature for 15 mins. Cells were then probed with ab22550 at 1/100 for 1 hour at room temperature. The secondary used was a DyLight® 488 goat anti-mouse used at 1/400 for 30 minutes at room temperature. Additional counterstains used were F-actin with a DyLight® 554 Phalloidin and Neuclei stained using a Hoechst 33342 conjugate. Image was taken at X20 magnification.

  • Chromatin Co-Immunoprecipitation (ChIP) analysis  using ab22550 binding acetylated lysines in 10E+06 LNCaP cells. Protein binding was detected using real-time PCR.

    Positive control: Fold enrichment of ab22550.
    Negative Control: Non-specific IgG.

  • ICC/IF image of ab22550 stained MCF7 cells. The cells were 4% formaldehye fixed (10 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 (ab22550, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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.

Anti-acetyl Lysine antibody [1C6] - ChIP Grade (ab22550)参考文献

This product has been referenced in:
  • Stice JP  et al. 17{beta}-Estradiol, Aging, Inflammation, and the Stress Response in the Female Heart. Endocrinology 152:1589-98 (2011). WB ; Rat . Read more (PubMed: 21303943) »

See 1 Publication for this product

Product Wall

Application ChIP
Detection step Real-time PCR
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Negative control mouse IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 8 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Positive control H2AZ
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提交于 Dec 15 2014

Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Total protein in input 20 µg
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提交于 Dec 15 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Denaturing (15)
Sample Mouse Cell lysate - whole cell (Hepatocyte)
Specification Hepatocyte
Blocking step I-Block(Applied biosystems) as blocking agent for 30 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
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提交于 Mar 11 2014

Thank you very much for your interest in ab22550. To our knowledge, ab22550 has not been tested in IP. Therefore, I can offer a discount off a future purchase if you buy ab22550 now, test it in IPand submit feedback to us in the form of an Abreview. It...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Jurkat cells)
Loading amount 10 µg
Specification Jurkat cells
Treatment trichostatin A 250 nM treated cells for 24h
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 May 19 2010

Thank you for your enquiry. The synthetic peptide used to raise this antibody was a peptide sequence surrounding the acetylated lysine 9 of histone H3. This antibody has been shown to generate a ladder in a HeLa whole cell lysate demonstrating that...

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Thank you for your enquiry. Yes, by Western blot, this antibody detects a ladder of acetylated proteins in HeLa cell lysates. In immunofluorescence procedures, this antibody recognizes acetylated lysine in gamma irradiated HeLa cells. I hope this...

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