Support FC. Each support was deprotected with AMA or ammonium hydroxide. The supernatant was removed and transferred to a cuvette and fesh solution was added to the support. The total exposure time of each sample is shown. Figure 3: dePrOteCtiOn and dePhOsPhOrylatiOn Of glen unysuPPOrt and glen unysuPPOrt fC
Solution Phase:
Ammonium Hydroxide:MethylAmine (AMA) 1:1 for 1 hour at 65 or Ammonium Hydroxide for 8 hours at 55. For sensitive minor bases or dyes, Glen UnySupport may be eliminated with 50 mM Potassium Carbonate in Methanol in 17 hours at room temperature or with Tert-Butylamine/Water 1:3 (v/v) for 4 hours at 60.
Gas Phase:
Methylamine gas for 30 minutes at 65 at 30 psi.
11
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We conclude that the minimum cleavage times are as follows: AMA NH4OH Glen UnySupport 10 min. 40 min. Glen UnySupport FC 2 min. 5 min. orderinG inForMation
Item
and the solution pushed back and forth until the next time point was reached. The total exposure time of each sample is shown in the illustration on Page 11. For Glen UnySupport, the solution after 10 minutes exposure to AMA is still pale blue indicating that a minimum of 10 minutes is required for cleavage. Similarly, with ammonium hydroxide, the solution after a total of 40 minutes exposure is still pale blue and so that would be the minimum time required for cleavage. In the same experiment with Glen UnySupport FC, the sample using AMA cleavage was pale blue after a total of only 2 minutes exposure.134-04-3 References More surprisingly, the cleavage was essentially complete in 5 minutes using ammonium hydroxide.1264-72-8 manufacturer It has to be stressed that these reactions are simply cleavage from the support. The oligos must still be fully deprotected, including dephosphorylation before use.
We are delighted to offer Glen UnySupport FC attached to 1000CPG in a variety of formats suited to high throughput synthesis, as well as in bulk for more routine use.
These products are covered by US Patent 7,202,264 owned by Isis Pharmaceuticals, Inc.
dNa CONjUgatiON tO HaLOtaggEd pROtEiN UsiNg gLEN BROMOHExyL LiNkER
Vijay Singh and Eric T. Kool Department of Chemistry Stanford University Stanford, CA 94305 (USA) The strategy of small-molecule fluorescent labeling of genetically encoded proteins has become a popular alternative to GFP labeling. This is because the methodology offers the advantage of much greater temporal resolution as the dye can be added at any point during the cell cycle or stage of an organism’s development. Among the most widely used approaches is the HaloTag method developed by Promega, which utilizes a bacterial haloalkane dehalogenase.PMID:25905177 The enzyme removes halides from aliphatic hydrocarbons by a nucleophilic displacement mechanism to form a covalent ester linkage between the haloalkane and Asp106 in the enzyme. In the wild type haloalkane dehalogenase, the ester is quickly hydrolyzed by histidine 272 in the catalytic active site. However, by mutating the histidine to phenylalanine, the HaloTag variant renders the covalent ester bond stable toward hydrolysis, as shown in Figure 1. The HaloTag domain is an exceedingly simple
Crystal struCture of Haloalkane DeHalogenase
VOLUME 26 NUMBER 1 May 2014
ac-5-Me-dC aMa REViEw CLiCk UpdatE tHpta
way to form covalent bonds between a variety of different molecules and labels to essentially any protein of interest.1 Standard ligands for HaloTag protein use lengthy chloroalkyl-substituted groups. Previously, we described a.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com