MaxLight™550 is a new Yellow-Green photostable dye conjugate comparable to Alexa Fluor™546, 555, DyLight™549 , Cy3™, TRITC and offers better labeling efficiency, brighter imaging and increased immunodetection.
The covalent modification of histone tails has regulatory roles in various nuclear processes, such as control of transcription and mitotic chromosome condensation.
Among the different groups of enzymes known to catalyze the covalent modification, the most recent additions are the histone methyltransferases (HMTases), whose functions are now being characterized.
Here we show that a SET domain-containing protein, G9a, is a novel mammalian lysine-preferring HMTase.
Like Suv39 h1, the first identified lysine-preferring mammalian HMTase, G9a transfers methyl groups to the lysine residues of histone H3, but with a 10-20-fold higher activity.
It was reported that lysines 4, 9, and 27 in H3 are methylated in mammalian cells.
G9a was able to add methyl groups to lysine 27 as well as 9 in H3, compared with Suv39 h1, which was only able to methylate lysine 9.
Our data clearly demonstrated that G9a has an enzymatic nature distinct from Suv39 h1 and its homologue h2.
Finally, fluorescent protein-labeled G9a was shown to be localized in the nucleus but not in the repressive chromatin domains of centromeric loci, in which Suv39 h1 family proteins were localized.
This finding indicates that G9a may contribute to the organization of the higher order chromatin structure of non-centromeric loci.
Applications:Suitable for use in Immunoblot.
Other applications not tested.
Recommended Dilution:Western Blot: 0.2-2ug/ml detected G9a in HeLa nuclear extract cell lysate.
HeLa nuclear extract was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-G9a (0.5ug/ml).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Optimal dilutions to be determined by the researcher.
Included Positive Antigen Control:HeLa nuclear extract.
Add an equal volume of Laemmli reducing sample buffer to 10ul of extract and boil for 5 minutes to reduce the preparation.
Load 20ug of reduced extract per lane for minigels.
Storage and Stability:Store product at 4°C in the dark.
DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid.
Dilute required amount only prior to immediate use.
Further dilutions can be made in assay buffer.
Caution: MaxLight™550 conjugates are sensitive to light.
For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Note: Applications are based on unconjugated antibody.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein A affinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™550.
Specificity：Recognizes human G9a protein, Mr ~140kD.
Isotype：IgG
Calc Applications Abbrev：WB
Calc Crossreactivity：Hu
Immunogen：KLH-conjugated, synthetic peptide (CDERVDSDSKSEVEALTEQ)corresponding to aa71-88 of human G9a, with an N-terminalcysteine added for conjugation purposes. Species sequence homology: mouse (17/18)
