Alkaline phosphatase conjugation

An enzyme-amplified detection scheme, based on tire coupling of a streptavidin-alkaline phosphatase conjugate and biotinylated target sequences was then applied. The enzyme catalysed the hydrolysis of the elecn oiiractive a-naphthyl phosphate to a-naphtlrol this product is elecU oactive and has been detected by means of differential... [Pg.15]

Figure 3 Immuno localization of acetyl esterase. Sections were incubated with antibodies raised against the acetyl esterase, followed by visualization with alkaline phosphatase conjugated secondary antibodies and staining with Fast Red.

Dendrimer-Alkaline Phosphatase Conjugate via thioether linkage... [Pg.375]

Alkaline Phosphatase Conjugation to Cystamine-Modified DNA Using Amine- and Sulfhydryl-Reactive Heterobifunctional Crosslinkers... [Pg.993]

Alkaline Phosphatase Conjugation to Diamine-Modified DNA Using DSS... [Pg.994]

Farmer, J.C., and Castaneda, M. (1991) An improved preparation and purification of oligonucleotide-alkaline phosphatase conjugates. Bio. Tech. 11, 588-589. [Pg.1062]

Jablonski, E., Moomaw, E.W., Tullis, R.H., and Ruth, J.L. (1986) Preparation of oligo-deoxynucle-otide-alkaline phosphatase conjugates and their use as hybridization probes. Nucleic Acids Res. 14, 6115-6129. [Pg.1078]

Anti-tumor effects of antibody-alkaline phosphatase conjugates in combination with etoposide phosphate. Proc. Natl. Acad. Sci. USA 85, 4842. [Pg.1112]

Note Do not use PBS for alkaline phosphatase-conjugated antibodies, since phosphate is an inhibitor of alkaline phosphatase. [Pg.26]

The Abbott IMx , a dedicated commercial immunoassay analyzer that employs FPIAs for small molecules, can also determine larger analytes by a fluorescence-based microparticle capture enzyme immunoassay (MEIA).(44) In this system, antibody-coated0.47- mlatexparticles are used for both sandwich and competitive assays, and alkaline phosphatase conjugates that bind to the particles cleave 4-methylumbelliferyl phosphate to generate the fluorophore. [Pg.465]

Figure 14.8. Enzymatically-amplified TR-FIA ofa-fetoprotein (AFP), using streptavidin-alkaline phosphatase conjugate (SA-ALP) and 5-lluorosalicyl phosphate (FSAP) to generate 5 fluorosalicylic acid (FSA). The FSA comhines with terbium-EDTA to form a bright, fluorescent complex. (Adapted from Ref. 83 with permission.) (01992, American Chemical Society).

Sodium azide is a powerful inhibitor of HRP, but sodium azide can be used with alkaline phosphatase conjugated antibodies without harmful effects. In addition, tap water or water deionized with polystyrene resins may inactivate the enzyme conjugate. Only use distilled, deionized water Tween-20 may interfere with some antibody-antibody reactions or may wash the protein of interest off the membrane. Tween-20 may be left out of the washes, but this may result in increased background... [Pg.213]

This protocol describes the use of an alkaline phosphatase conjugate of course, a HRP conjugate works well, too. [Pg.160]

Incubate with the alkaline phosphatase-conjugated secondary antibody (30 min)... [Pg.113]

Fig. 1. Double label immunohistochemistry on rai liver. An acetone-fixed rat liver section was incubated with a polyclonal antiserum raised in rabbit to a hepa-tocyte cell surface protein (courtesy of Dr. S. Stamatoglou) and a mouse monoclonal antibody against a bile duct specific cytokeratin (courtesy of Dr. E. B. Lane). The hepatocyte protein was localized by use of a secondary peroxidase-conjugated antibody resulting in a red/brown product (thin arrow). The bile duct cytokeratin was identified by using an alkaline phosphatase-conjugated secondary antibody giving a blue color (thick arrow).

Alkaline phosphatase-conjugated antimouse IgG raised in goat. [Pg.272]

Incubate the slides m alkaline phosphatase-conjugated antidigoxigemn diluted 1.600 in TBT. [Pg.393]

Incubate the slides m a mixture of avidin—peroxidase conjugate diluted 1.100 and alkaline phosphatase-conjugated antidigoxigenin diluted 1 600 m TBT, in practice, 1 pL of antibody and 6 pL of avidin are added to 600 pL of TBT. [Pg.393]

Nucleic acids can also be biotinylated by nonenzymatic methods with photobiotin, a photoactivatable biotin analog (6), which can be commercially obtained from BRL, Sigma, and other commercial sources I have not compared the suitability of this method of biotin incorporation with that reported here, but expect that the method would be fully acceptable FMC (Rockland, ME) markets an alternate nonradioactive sequence detection kit known as Chemiprobe. The basis of this system is a chemical modification of cytosine residues m the probe DNA. After hybridization, the probe is detected by means of a monoclonal antibody that specifically recognizes the sulfonated DNA. Detection of the bound monoclonal antibody is achieved by means of an alkaline phosphatase-conjugated second antibody. [Pg.403]

Dilute alkaline phosphatase conjugated rabbit antigoat IgG 1 1000 in PBST and dispense 200 pL/well Incubate at 37°C for 1 h... [Pg.435]

Add the reveal antiserum (for example, rabbit antigoat IgG alkaline phosphatase conjugate) at an appropriate dilution (1-1000 in Tris/saline), and incubate for 1 h at room temperature. [Pg.436]

Add secondary antibodies (as for ELISA and Western blotting, but use an alkaline-phosphatase conjugate instead of a HRP) in TBS/Tween-20/BSA Incubate 30 min at room temperature... [Pg.495]

Blake, M.S., Johnston, K.H., Russell-Jones, G.J., and Gotschlich, E.C. 1984. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal. Biochem. 136 175-179. [Pg.217]

Senter, P. D., Saulnier, M. G., Schreiber, G. J., Hirschberg, D. L., Brown, J. P., Hellstrom, I., and Hellstrom, K. E. (1988) Anti-tumor effects of antibody—alkaline phosphatase conjugates in combination with etoposide phosphate. Proc. Natl. Acad. Sci. U.S.A. 85, 4842. [Pg.734]

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