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In Situ Hybridization and Detection Reagents
RNA Probe Design and Generation – How are RNA probes prepared?
RNA probes are usually prepared with biotin or specific fluorophore by in vitro transcription. High specific activity RNA probes or riboprobes…
Tue, Jul 18, 2023
How are ISH probes labeled?
There are three main techniques for probe labeling. Labeled probes can be generated by incorporating labeled nucleotides during the synthesis…
Tue, Jul 18, 2023
What are the advantages of digoxigenin labeling?
For chromogenic ISH, Digoxigenin, compared to biotin, offers the advantage of lower background, higher specificity. Is also possible to combine…
Tue, Jul 18, 2023
Why should post-hybridization washes be carried out?
Post-hybridization washing after ISH will reduce non-specific signal by removing probes that are lightly bound to off-target sequences. Most frequently,…
Tue, Jul 18, 2023
How can slides be stored after ISH?
That depends mostly on fixation/embedding method and the type of ISH performed. After chromogenic ISH on FFPE or cryopreserved tissue,…
Tue, Jul 18, 2023
How big are ISH probes?
There is no defined answer. Depending on the tissue specimen, target type, and sequence, ISH probes might vary from below…
Tue, Jul 18, 2023
Probe Design and Generation – How are DNA probes prepared?
First choice is to explore if commercially ready-to-use probes are available for specific targets. Enzo Life Sciences offers a selection…
Tue, Jul 18, 2023
How can RNA degradation in the sample be prevented?
When RNA is the target of ISH, preservation of RNA in the sample is very important given the ubiquitous presence…
Tue, Jul 18, 2023
Is there a specifc type of glass slide recommended for ISH?
Positively charged glass slides have proven optimal to draw tissue sample sections for subsequent ISH on. Alternatively, poly-L-lysine or silanized…
Tue, Jul 18, 2023
Tissue specimen preparation – How should samples be stored and for how long?
While whole tissue specimen are sometimes used for long-term storage, most often glass slide-mounted tissue sections are stored at 4°C.…
Tue, Jul 18, 2023