Biosearch Technologies
Stellaris RNA FISH
VSMF-1015-5
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The overall objective of this objective is to understand host cell responses to the infection. The specific goal is to phenotype the cells that get infected during the course of HIV/SIV infection. Stellaris FISH probes were chosen because there are no antibodies available for the transcription factor we were interested in and we wanted to see what percentage of specific cell type was expressing this specific transcription factor.
Immunofluorescence
Rhesus Macaque rectal tissue
20-micron thick tissue sections were fixed with 3.7% (vol./vol.) formaldehyde in 1X PBS, then blocked with NDS and stained with antibodies of choice. Prior to FISH staining the slides were incubated with 70% EtOH for 10 minutes at the room temperature. The remainder of RNA-FISH staining was performed according to the manufacturer’s recommendations for sequential IF and FISH procedures (Biosearch Technologies, CA). In brief slides were incubated in Stellaris FISH blocking reagent for 30 minutes and were than incubated with RNA probe (1:1000 dilution) for 1 hour at room temperature.
Take all attention to minimize the exposure of tissue and reagents to RNases.
We were able to perform combination of immunofluorescence staining with RNA Fluorescent in situ hybridization. We were able to observe distinct staining patterns of our antibodies and RNA probe and demonstrated that large percentage of our cell type expressed the transcription factor we were interested in. The probe signal was very bright and punctuate in appearance.
None
Very bright and high specificity
Price
Great way to stain for the targets to which antibodies are not available.
Product Name: Stellaris® FISH Probes, eGFP with Quasar® 670 DyeSupplier Page