Acetyl-Lysine Antibody (1C6)

Western Blot: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Analysis of Acetyl-Lysine in HeLA cells.

Immunocytochemistry/Immunofluorescence: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Analysis of lysine acetylated proteins (green) in HeLa cells left untreated (left panels) or treated with 0.3uM Trichostatin A for 16 hours (right panels).

Chromatin Immunoprecipitation: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Analysis of chromatin proteins with Acetyl-Lysine was performed using cross-linked chromatin from 4-10^6 LNCaP cells. Immunoprecipitation was performed using the Magnetic ChIP kit with 10ug of an Acetyl-Lysine monoclonal antibody. Quantitation of immunoprecipitated GAPDH promoter sequence is presented as fold enrichment of the acetyl lysine monoclonal antibody versus non-specific IgG.

Western Blot: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Analysis of Acetyl-Lysine was performed by loading various amounts of acetylated BSA or non-acetylated BSA per well and 10ul of PageRuler Prestained Protein Ladder onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Acetyl-Lysine monoclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico.

Western Blot: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Analysis of lysine acetylated proteins from cells left untreated (DMSO only) or cells treated with 0.3uM or 3uM of Trichostatin A (TSA) for 16 hours was performed by loading 50 ug of the indicated whole cell lysates per well and 10ul of PageRuler Prestained Protein Ladder onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Acetyl-Lysine monoclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico.

Dot Blot: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Specific detection of lysine acetylated histones using an Acetyl-Lysine monoclonal antibody. Histone peptides containing 384 different modification combinations spotted in duplicate on a peptide array were used according to the manufacturer instructions. A schematic and table with representative results (positive detection in red, negative in blue) are shown. Result interpretation and quantification were performed using the manufacturer software. Chemiluminescent detection was performed.

Immunoprecipitation: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Immunoprecipitation of acetylated alpha-Tubulin was performed.3uM or 3uM Trichostatin A (TSA) for 16 hours. Antigen-antibody complexes were formed by incubating 500ug of lysate with 3ug of an Acetyl-Lysine monoclonal antibody overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Agarose at a dilution of 1:1000 overnight rotating at 4C, washed in TBST, and probed with Clean-blot IP Detection Reagent at a dilution of 1:2000 for at least 1 hour.

Immunoprecipitation: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Analysis of acetylated Histone H3 (Lys9) was performed using whole cell lysates from cells left untreated (DMSO only) or cells treated with 0.3uM or 3uM Trichostatin A (TSA) for 16 hours. Antigen-antibody complexes were formed by incubating 500ug of the indicated lysate with 3ug of an Acetyl-Lysine mAb overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Agarose, washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hr. The membrane was probed with an Acetyl-Histone H3 (Lys9) mAb at a dilution of 1:1000 overnight rotating at 4C, washed in TBST, and probed with Clean-blot IP Detection Reagent at a dilution of 1:2000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico.

Dot Blot: Acetyl-Lysine Antibody (1C6) [NB100-74339] - Specific detection of lysine acetylated histones using an Acetyl-Lysine monoclonal antibody. Histone peptides containing 384 different modification combinations spotted in duplicate on a peptide array were used according to the manufacturer's instructions (Active Motif). Representative results showing specific detection of acetylated Histone H4 K20 (blue circles, panel A) and H3 K9 (blue circles, panel B). Note that Histone H4 K20me2/K20me3 (red and yellow circles, panel A) and H3 K9me2/K9m3 (red and yellow circles, panel B) are not detected by the Acetyl-Lysine monoclonal antibody.

• Reactivity: All-NA
• Applications: WB, DB, ICC/IF, IP, ChIP
• Clone: 1C6
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Acetyl-Lysine Antibody (1C6) Summary

• Immunogen: Synthetic peptide sequence surrounding the acetylated lysine 9 of histone H3.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 10 ug
  • Dot Blot 1:100 - 1:2000
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:500
  • Immunoprecipitation 3 ug
  • Western Blot 1:500 - 1:2000

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS, 1 mg/mL BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for Acetyl-Lysine Antibody (1C6)

• acetyl Lysine
• AC-LYS-OH
• Lysine
• N-acetyl lysine

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Acetyl-Lysine Antibody (NB100-74339)(2)

Publications using NB100-74339

• Dziewulska A, Dobosz AM, Dobrzyn A et al. SCD1 regulates the AMPK/SIRT1 pathway and histone acetylation through changes in adenine nucleotide metabolism in skeletal muscle J. Cell. Physiol. 2019-06-26 [PMID: 31241768] (Mouse)
• Tataranni T, Agriesti F, Ruggieri V et al. Rewiring carbohydrate catabolism differentially affects survival of pancreatic cancer cell lines with diverse metabolic profiles Oncotarget 2017-06-20 [PMID: 28476035] (WB, Human)

Reviews for Acetyl-Lysine Antibody (NB100-74339) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Mouse.

reviewed by: Verified Customer ICC Mouse 11/05/2021

Summary

• Application: Immunocytochemistry
• Sample Tested: Hepatic stellate cells
• Species: Mouse
• Lot: 1C6

Comments

• Comments: Primary antibody dilution 1:100

reviewed by: Michaela Rekow WB Mouse 01/14/2016

Summary

• Application: Western Blot
• Sample Tested: Splenocytes
• Species: Mouse