Navigation Links
Detection of mRNAs on Cryosections of the,,,Cardiovascular System Using DIG-Labeled RNA Probes

g solution: 335 g NBT [stock solution: 75 mg/ml in 70 % dimethylformamide], 174 g BCIP [stock solution: 50 mg/ml in 100% dimethylformamide], and 240 g Levamisole per ml buffer 3). The staining solution was removed by rinsing in 5x TE, the staining procedure was stopped by incubating the slides for 15 minutes in 5x TE. The sections were briefly rinsed in distilled water, then counterstained with methylene green. The sections were mounted with Kaisers glycerin-gelatin. If immunofluorescence protocols are used in combination with in situ hybridization, we recommend development with the ELF substrate (ELF Kit, Molecular Probes).

  • Fluorochrome-conjugated antibodies

When the mRNA of interest is abundantly expressed, anti-DIG antibodies conjugated with FITC or other fluorochromes can be used. The slides were incubated for 2 hours with FITC-conjugated anti-DIG antibody (1:201:200 in buffer 1), then washed twice for 5 minutes with buffer 2 (Buffer 1 containing 0.05% Tween). The sections were counterstained with Hoechst Dye 33342 and mounted with fluorescence mounting medium (DAKO).

Immunohistochemistry

Usually immunohistochemistry was performed immediately after in situ hybridization. However, with both detection procedures excellent results were also obtained for several weeks (up to months) after in situ hybridization. All washing steps were performed on a shaking platform.

  • The peroxidase staining procedure was performed according to the manufacturers recommendations (Vectastain Elite Kit, Vector Laboratories).
  • Immunofluorescence: The slides were incubated with PBS/12% BSA for 12 hours in a humidified chamber, then the blocking solution was removed. The slides were incubated with an appr
    '"/>


Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A New PCR-based Mycoplasma Detection Method
3. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
4. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
5. Gene Expression Arrays: Highly Sensitive Detection of Expression Patterns with Improved Tools for Target Amplification
6. The DIG System Nonradioactive and Highly Sensitive Detection of Nucleic Acids
7. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
8. Quantification of Nucleosomes in Serum by the Cell Death Detection ELISAplus
9. A Further Step in Understanding Apoptosis Direct Detection of PARP Cleavage
10. In Situ Cell Death Detection Kit
11. Detection of antibody-stained cell surface and intracellular protein targets with the Agilent 2100 bioanalyzer
Post Your Comments:
(Date:10/1/2014)... 30, 2014 In response to ... to address sales, training and consumer engagement challenges, ... innovative mobile applications to turn their clients’ visions ... latest customized mobile app creations, are now featured ... In partnership with Toyota, StudioPMG created a robust ...
(Date:10/1/2014)... (PRWEB) October 01, 2014 The U.S. ... to Southern Illinois University Edwardsville as part of the ... initiative. The funding is co-administered by the Department of ... Biden, Secretary of Labor Thomas E. Perez and Secretary ... million in job-driven training grants to nearly 270 colleges ...
(Date:9/30/2014)... German . ... have demonstrated a new kind of building block for ... chips could be based on three-dimensional arrangements of nanometer-scale ... of the semiconductor industry CMOS fabrication of silicon ... collaborators at the University of Notre Dame are exploring ...
(Date:9/30/2014)... Sept. 30, 2014 Nematicide is a ... Nematodes are microscopic parasitic roundworms, found in massive ... other plants and animals. Nematicides have tended to ... properties promoting migration through the soil. A single ... plant-parasitic nematodes. With more than 10,000 species classified, ...
Breaking Biology Technology:Digital Agency Studio PMG Launches New Case Studies on Website 2U.S. Dept. of Labor Awards SIUE Nearly $10-Million Grant 2U.S. Dept. of Labor Awards SIUE Nearly $10-Million Grant 3U.S. Dept. of Labor Awards SIUE Nearly $10-Million Grant 4A new dimension for integrated circuits: 3-D nanomagnetic logic 2A new dimension for integrated circuits: 3-D nanomagnetic logic 3Nematicides Market by Types (Fumigants, Organophosphates, Carbamate and Bio-Nematicides), Applications (Agrochemicals, Industrial, and Others), and Geography (North America, Europe, Asia-Pacific & ROW) - Global Trends & Forecasts to 2019 2
... , SAN MATEO, Calif., Aug. 25 NeurogesX, Inc. ... and commercializing novel pain management therapies, today announced the appointment of ... and Audit Committee chairman, effective August 20, 2009. Mr. Goodwin currently ... on exploiting a novel, Nobel Prize winning method of gene silencing. ...
... , GAITHERSBURG, Md., Aug. 25 GenVec, Inc. (Nasdaq: ... Phase 2 Small Business Innovation and Research (SBIR) grant from the ... Institutes of Health (NIH) to support the development of GenVec,s vector ... http://www.newscom.com/cgi-bin/prnh/ 2 0081205/DC50112LOGO ) , , ...
... , , , COVINA, Calif., ... banks, today announced that Shannon Miller , a 7-time Olympic Medalist ... appear in a series of advertising and public awareness campaigns for the ... , , Considered "The Most Decorated American Gymnast" by ...
Cached Biology Technology:NeurogesX Appoints New Board Director and Audit Committee Chairman 2NeurogesX Appoints New Board Director and Audit Committee Chairman 3NeurogesX Appoints New Board Director and Audit Committee Chairman 4NeurogesX Appoints New Board Director and Audit Committee Chairman 5GenVec Receives an Approximately $2.5 Million Grant to Support Novel Cell Line Development 27-Time Olympic Medalist Shannon Miller Endorses StemCyte 2
(Date:10/1/2014)... the deserts, rainforests, prairies and forests of the world ... Central Park, according to a surprising new study led ... Boulder. , The research team analyzed 596 soil ... discovered a stunning diversity of below-ground life, most of ... 8.5 percent to 16.2 percent of the organisms discovered ...
(Date:9/30/2014)... the University of California, San Diego, have completed the ... a method that uses the pixels in an image ... as mesh-free simulation. , The researchers, led by J.S. ... the Jacobs School of Engineering at UC San Diego, ... at the CompIMAGE,14 conference in Pittsburgh this month. Chen ...
(Date:9/30/2014)... have in common? Unlike most eukaryotic multicellular organisms ... these organisms are all polyploid, meaning they have three ... 3 and 4 sets of chromosomes, respectively, and strawberries ... fact most plant species are polyploid. Polyploidy, or genome ... only recently, with the development of molecular tools, has ...
Breaking Biology News(10 mins):Microbes in Central Park soil: If they can make it there, they can make it anywhere 2Microbes in Central Park soil: If they can make it there, they can make it anywhere 3First comprehensive meshfree numerical simulation of skeletal muscle tissue achieved 2Gene doubling shapes the world: Instant speciation, biodiversity, and the root of our existence 2Gene doubling shapes the world: Instant speciation, biodiversity, and the root of our existence 3Gene doubling shapes the world: Instant speciation, biodiversity, and the root of our existence 4
... new research shows that, contrary to conventional belief, ... polysaccharideare present in abundance in fossils of arthropods ... signature of chitin-protein complex was discovered in 25 ... structural protein have also been discovered in 80 ...
... Women who are satisfied with everyday life and are ... reveals a thesis from the University of Gothenburg, Sweden. ... has looked at how everyday life affects alcohol consumption ... Gothenburg (WAG) population study which has been ongoing at ...
... sources of soybean meal are capturing attention throughout the country. ... enzyme-treated soybean meal may replace fish meal in weanling pig ... is causing producers to search for new options for weanling ... of animal sciences. "Pigs are traditionally fed diets containing relatively ...
Cached Biology News:Unexpected exoskeleton remnants found in Paleozoic fossils 2Women involved in leisure activities drink less alcohol 2Research proves new soybean meal sources are good fish meal alternatives 2
... than western blotting. Get results sooner and save ... gel detection. Eliminates the need for a membrane ... All reagents are ready-to-use. No reagents to prepare ... 2 reagents required. Guarantees a simple to perform, ...
... Plasmid-Based Deletion Machine is based on the ... developed by Goryshin and Reznikoff, and by ... families of deletions and inversions by intramolecular ... transposon-containing plasmid vector. Although most researchers will ...
... This kit is designed to knockout or ... in less than one week. Red/ET recombination allows ... pair precise, specific, and faithful manner. An FRT-flanked ... kit which can be used to replace a ...
5X StabilCoat Immunoassay Stabilizer is a 5X concentrate solution of StabilCoat Immunoassay Stabilizer. The shelf-life of 5X StabilCoat Immunoassay Stabilizer is 2 years when stored at 4C. Product ...
Biology Products: