Anti-acetyl Lysine antibody - ChIP Grade (ab21623)
Key features and details
- Rabbit polyclonal to acetyl Lysine - ChIP Grade
- Suitable for: IP, IHC-P, ChIP, ICC/IF, WB, ELISA
- Reacts with: Species independent
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
-
Product name
Anti-acetyl Lysine antibody - ChIP Grade
See all acetyl Lysine primary antibodies -
Description
Rabbit polyclonal to acetyl Lysine - ChIP Grade -
Host species
Rabbit -
Specificity
Recognises proteins acetylated on lysine residues. Tested: acetylated histone, acetylated BSA, and acetylated MBP, no reaction to the non acetylated proteins. -
Tested applications
Suitable for: IP, IHC-P, ChIP, ICC/IF, WB, ELISAmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule corresponding to acetyl Lysine conjugated to keyhole limpet haemocyanin. Acetylated KLH conjugates.
-
General notes
Use to detect acetylation of lysine.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 6.00
Preservative: 0.1% Sodium azide
Constituent: Tris buffered saline -
Concentration information loading...
-
Purity
Immunogen affinity purified -
Purification notes
The antibody was specifically purified with immobilised acetylated lysine on agarose -
Primary antibody notes
Use to detect acetylation of lysine. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
-
ChIP Related Products
-
Compatible Secondaries
-
Isotype control
Applications
The Abpromise guarantee
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 | (1) |
Use at an assay dependent concentration.
|
IHC-P |
Use at an assay dependent concentration. PubMed: 24160175
|
|
ChIP | (1) |
Use 2-4 µg for 25 µg of chromatin.
|
ICC/IF | (1) |
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 | (7) |
1/1000 - 1/2500.
|
ELISA |
1/5000.
|
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
-
Relevance
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. -
Alternative names
- pan acetyl Lysine antibody
Images
-
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.
-
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 -
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
All lanes : 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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
References (61)
ab21623 has been referenced in 61 publications.
- Zhao J et al. Mitochondrial PKM2 deacetylation by procyanidin B2-induced SIRT3 upregulation alleviates lung ischemia/reperfusion injury. Cell Death Dis 13:594 (2022). PubMed: 35821123
- Zhang J et al. Adiponectin ameliorates hypertrophic scar by inhibiting Yes-associated protein transcription through SIRT1-mediated deacetylation of C/EBPβ and histone H3. iScience 25:105236 (2022). PubMed: 36274941
- Takakura M et al. Rpd3/CoRest-mediated activity-dependent transcription regulates the flexibility in memory updating in Drosophila. Nat Commun 12:628 (2021). PubMed: 33504795
- Jayakumari NR et al. Honokiol regulates mitochondrial substrate utilization and cellular fatty acid metabolism in diabetic mice heart. Eur J Pharmacol 896:173918 (2021). PubMed: 33529726
- Feng L et al. Sirt1 deacetylates and stabilizes p62 to promote hepato-carcinogenesis. Cell Death Dis 12:405 (2021). PubMed: 33854041