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Immunofluorescent 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). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with an Acetyl Lysine monoclonal antibody (Product # MA1-2021) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834). A merged image with nuclei staining (blue) using Hoechst 33342 dye (Product # 62249) is shown on bottom panels. Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Chemical|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||Synthetic peptide sequence surrounding the acetylated lysine 9 of histone H3.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||10 µg|
|Dot blot (DB)||Assay dependent|
|Immunocytochemistry (ICC)||1:100 - 1:500|
|Immunofluorescence (IF)||1:100 - 1:500|
|Immunoprecipitation (IP)||3 µg|
|Western Blot (WB)||1:500 - 1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
MA1-2021 detects human and mouse, and monkey acetylated lysine.
MA1-2021 has been used successfully in Western blot, immunoprecipitation, dot blot, ChIP Assay, and immunofluorescence (cells) procedures. By Western blot, this antibody may detect a ladder of acetylated proteins in HeLa cell lysate. In immunofluorescence procedures, MA1-2021 recognizes acetylated lysine in gamma irradiated HeLa cells.
The MA1-2021 immunogen is a synthetic peptide sequence surrounding the acetylated lysine 9 of histone H3.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
MFN1 deacetylation activates adaptive mitochondrial fusion and protects metabolically challenged mitochondria.
MA1-2021 was used in western blot to determine if mitochondrial fusion protects metabolically challenged mitochondria
|Lee JY,Kapur M,Li M,Choi MC,Choi S,Kim HJ,Kim I,Lee E,Taylor JP,Yao TP||Journal of cell science (127:4954)||2014|
ARD1 stabilization of TSC2 suppresses tumorigenesis through the mTOR signaling pathway.
MA1-2021 was used in western blot to assess whether ARD1 mediates epsilon-acetylation of TSC2
|Kuo HP,Lee DF,Chen CT,Liu M,Chou CK,Lee HJ,Du Y,Xie X,Wei Y,Xia W,Weihua Z,Yang JY,Yen CJ,Huang TH,Tan M,Xing G,Zhao Y,Lin CH,Tsai SF,Fidler IJ,Hung MC||Science signaling (3:null)||2010|