A rapid colorimetric in situ mRNA hybridization technique using hyperbiotinylated oligonucleotide probes for analysis of mdr1 in mouse colon carcinoma cells. 1993

C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
Department of Cell Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.

We describe the development of a rapid colorimetric in situ hybridization technique utilizing oligonucleotide probes labeled with six biotin molecules at the 3' end to detect mdr1 in mouse colon cancer cells growing in culture and in vivo. mRNA integrity was verified by the use of a multibiotinylated poly d(T) oligonucleotide, and the specificity of the reaction was confirmed by use of labeled sense and anti-sense probes in serial cryostat sections and cultured cells. The multiple biotin label produced a strong signal after a short hybridization time. Avidin-alkaline phosphatase detection and the capillary technology used in the Microprobe Accelerated System allowed completion of the procedure in less than 5 hr. Excellent correlations with the MDR phenotype of the cells, Northern blot analysis, and immunohistochemistry recommend this procedure for identifying cells that express the MDR phenotype in culture and in vivo.

UI MeSH Term Description Entries
D008297 Male Males
D008562 Membrane Glycoproteins Glycoproteins found on the membrane or surface of cells. Cell Surface Glycoproteins,Surface Glycoproteins,Cell Surface Glycoprotein,Membrane Glycoprotein,Surface Glycoprotein,Glycoprotein, Cell Surface,Glycoprotein, Membrane,Glycoprotein, Surface,Glycoproteins, Cell Surface,Glycoproteins, Membrane,Glycoproteins, Surface,Surface Glycoprotein, Cell,Surface Glycoproteins, Cell
D008807 Mice, Inbred BALB C An inbred strain of mouse that is widely used in IMMUNOLOGY studies and cancer research. BALB C Mice, Inbred,BALB C Mouse, Inbred,Inbred BALB C Mice,Inbred BALB C Mouse,Mice, BALB C,Mouse, BALB C,Mouse, Inbred BALB C,BALB C Mice,BALB C Mouse
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D009363 Neoplasm Proteins Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm. Proteins, Neoplasm
D010641 Phenotype The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment. Phenotypes
D003110 Colonic Neoplasms Tumors or cancer of the COLON. Cancer of Colon,Colon Adenocarcinoma,Colon Cancer,Cancer of the Colon,Colon Neoplasms,Colonic Cancer,Neoplasms, Colonic,Adenocarcinoma, Colon,Adenocarcinomas, Colon,Cancer, Colon,Cancer, Colonic,Cancers, Colon,Cancers, Colonic,Colon Adenocarcinomas,Colon Cancers,Colon Neoplasm,Colonic Cancers,Colonic Neoplasm,Neoplasm, Colon,Neoplasm, Colonic,Neoplasms, Colon
D004351 Drug Resistance Diminished or failed response of an organism, disease or tissue to the intended effectiveness of a chemical or drug. It should be differentiated from DRUG TOLERANCE which is the progressive diminution of the susceptibility of a human or animal to the effects of a drug, as a result of continued administration. Resistance, Drug
D005629 Frozen Sections Thinly cut sections of frozen tissue specimens prepared with a cryostat or freezing microtome. Frozen Section,Section, Frozen,Sections, Frozen
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

Related Publications

C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
November 1995, The American journal of pathology,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
January 1994, Annals of clinical and laboratory science,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
May 1995, British journal of haematology,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
January 2020, Methods in molecular biology (Clifton, N.J.),
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
January 1991, Journal of biochemical and biophysical methods,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
January 2002, International review of neurobiology,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
September 2000, International microbiology : the official journal of the Spanish Society for Microbiology,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
December 1990, Journal of cellular biochemistry,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
January 1985, Journal of cellular biochemistry,
C D Bucana, and R Radinsky, and Z Dong, and R Sanchez, and D J Brigati, and I J Fidler
January 2007, Advances in clinical chemistry,
Copied contents to your clipboard!