Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays.
AuthorsBobrow MN, Harris TD, Shaughnessy KJ, Litt GJ
JournalJ Immunol Methods
PubMed ID2558138
'A novel signal amplification method, catalyzed reporter deposition (CARD), and its application to immunoassays is described. The method involves utilizing an analyte-dependent reporter enzyme (ADRE) to catalyze the deposition of additional reporter on the surface in a solid-phase immunoassay. In the examples described, deposition of reporter is facilitated by using ... More
Fluorescence probe studies of the state of tropomyosin in reconstituted muscle thin filaments.
AuthorsIshii Y, Lehrer SS
JournalBiochemistry
PubMed ID3663633
'The monomer fluorescence of N-(1-pyrenyl)maleimide-labeled tropomyosin bound to F-actin (PTm-actin) increases when myosin subfragment 1 (S1) binds to actin and is half complete when only approximately 1 S1 is bound to 7 actin subunits [Ishii, Y., & Lehrer, S. S. (1985) Biochemistry 24, 6631-6638]. Similar studies of the binding of ... More
Fluorescence sandwich enzyme-linked immunosorbent assay for detecting human interleukin-2 receptors.
AuthorsHonda M, Nagao S, Yamamoto N, Tanaka Y, Tozawa H, Tokunaga T
JournalJ Immunol Methods
PubMed ID2453583
A very sensitive fluorescence sandwich enzyme-linked immunosorbent assay (FS-ELISA) for the detection of soluble and cell-associated human interleukin-2 receptors (IL-2R) has been developed. For use in this assay system, two anti-human IL-2R monoclonal antibodies, H 48 and biotinylated HA 26, were selected from six monoclonal antibodies directed against different epitopes ... More
Measurement of antigen-specific mouse IgE by a fluorometric reverse (IgE-capture) ELISA.
AuthorsSakaguchi M, Inouye S, Miyazawa H, Tamura S
JournalJ Immunol Methods
PubMed ID2783445
A reverse, or IgE-capture, enzyme-linked immunosorbent assay (ELISA) for measuring ovalbumin-specific IgE antibody in the serum of immunized mice has been developed. Microplate wells were first coated with a commercial anti-mouse IgE rat monoclonal antibody, and then incubated with two-fold serial dilutions of test sera with 10% normal mouse serum ... More
TNF-alpha pretreatment prevents subsequent activation of cultured brain cells with TNF-alpha and hypoxia via ceramide.
AuthorsGinis I, Schweizer U, Brenner M, Liu J, Azzam N, Spatz M, Hallenbeck JM
JournalAm J Physiol
PubMed ID10329967
We have developed a cellular model in which cultured astrocytes and brain capillary endothelial cells preconditioned with tumor necrosis factor-alpha (TNF-alpha) fail to upregulate intercellular adhesion molecule-1 (ICAM-1) protein (80% inhibition) and mRNA (30% inhibition) when challenged with TNF-alpha or exposed to hypoxia. Inasmuch as ceramide is known to mediate ... More