Labeling in vitro

Adams SR, Campbell RE, Gross LA et al (2002) New Biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo synthesis and biological applications. J Am Chem Soc 124 6063-6076... 

Product label research use Product label investigational use Product label in vitro diagnostic use... 

Labeling in vitro may be also natural. Suitable methods have been developed in last ten years. 

Foreign labeling in vitro is certainly very widely used. The most important labeling methods which have been applied for labeling almost all important proteins are halogenation (iodination or bromation) and conjugation (reductive methylation) alkylation and acylation 3 )... 

Other possibilities are the radioactive labeling in vitro of one of the monoclonal antibodies and enzyme-labeling of the other and their detection in cells by autoradiography (Tijssen and Kurstak, 1977) or the complexing of heavy metals to one of the immunoreact-ants (e.g., colloidal gold to SpA Roth, 1982). 

Fig. 2 RBL-2H3 cell labeled in vitro with mAb AA4 directly conjugated to Texas Red after fixation. The plasma membrane is labeled by the antibody. Microvilli (arrow) can be seen on the cell surface.

Pectic acid has been esterified with propionic anhydride and the product partially reduced with gaseous diborane. Tritium labelling has been applied to heparin by using tritium gas under an electric discharge or tritiated water in the presence of palladium black. Labelled glycosaminoglycans may also be prepared by labelling in vitro. - ... 

Fig, 5 Isokinetic binding sedimentation of T3 receptors from oligodendrocytes and astrocytes. The receptors were extracted from nuclei of rat oligodendroglial and astroglial cells after 30 and 24 days in culture, respectively, with 0.4 M KCl and labelled in vitro with L-[ I]T3 in the absence ( -— ) or presence (O—O) of an excess of non-radioactive T3. The arrows represent the position of hemoglobin which has a sedimentation coefficient 4 3 S and was used as an internal marker (from Yusta et al.. Endocrinology 122 in press (1988) (71). With permission). 

A protocol for proteasome labeling in vitro and in situ with broad-spectrum biotin-or Bodipy-containing peptide vinyl sulfones or peptide epoxyketones. 

Figure 4 Overview of MS-based strategies for quantitative proteomics. Depending on the point at which the label is introduced, most procedures are classified as (A) in vivo labeling, (B) predigestion labeling in vitro, or (C) postdigestion labeling in vitro.

Nuclear and polysomal fractions were prepared from embryos labeled in vitro with [ HJuridine in the presence of actinomycin D at a concentration (0.04 /ig/ml) sufficient to inhibit rRNA synthesis as described by Schultz et al. (1973a). 

Early work performed by Borst and co-workers (Borst and Aaij, 1969 Aaij et ah, 1970) revealed that mt RNA radioactively labeled in vivo with P predominantly hybridizes to the H strand of the mt DNA (Table VIII), as does mt RNA labeled in vitro with [ H]nucleoside triphosphates. But in contrast to the mt RNA synthesized in vivo the product obtained in vitro to some extent also hybridizes with the L strand. 

If radioactive amino acids are incorporated into mitochondrial proteins in vitro radioactivity is associated with mitoribosomes (Ashwell and Work, 1970). But this activity can be removed by puromycin treatment, suggesting that the radioactivity is the result of incomplete peptide chains on the mitoribosomes. There is no evidence that any mitochondrial ribosomal proteins are labeled in vitro although a clear-cut separation of the ribosomal proteins from animal cells has not been reported. Even with N. crassa no evidence was found that ribosomal proteins are coded for by mt DNA (Lizardi and Luck, 1972). From all these data it seems very likely that mitochondrial proteins are synthesized outside the mitochondria, under control of the nuclear genome, and are subsequently transported into the mitochondrial compartment. 

BioArray High-Yield RNA Transcript Labeling System RNA Labeling In-Vitro Transcription Reaction... 

In the data shown in Fig. 1, soluble chromatin was applied to a poly(ADP-ribose) antibody column in PBS, and washed extensively. In earlier studies, nucleosomes were labeled in vitro with NAD prior to immunofractionation. In the experiments described [1-3], no in vitro synthesis of exogenous poly(ADP-ribosylation) was performed and absorption to the antibody column relied upon the presence of in vivo synthesized poly(ADP-ribose) at the sites in chromatin where it presumably exists naturally. This laboratory has recently shown the natural presence of 15 units of poly(ADP-ribose) attached to histone HI isolated from intact, unlabeled cells using the same antibody [5]. 

Analysis of Nuclear Acceptor Proteins for ADP-Ribose Labelled in Vitro... 

Binding Assay. 10 /ig sucrose-loaded liposomes were added to 10% of S-methionine labeled in vitro translated reaction product (5 /il) in 50 /il... 

Lang K, Davis L, Wallace S et al (2012) Genetic encoding of bicyclononynes and trans-cyclooctenes for site- specific protein labeling in vitro and in live mammalian cells via rapid fluorogenic Diels-Alder reactions. J Am Chem Soc 134 10317-10320... 

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