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Knowledge Base
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
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 are Aqua 431 dUTP-labeled probes much harder to detect?
There are two aspects making Aqua-labeled probes harder to detect: First, the aqua fluorophore (DEAC – 7-Diethylaminocoumarin-3-carboxylic acid) has a…
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
Can multiple targets be detected with ISH?
Yes, it is common practice to stain with multiple ISH probes in parallel. However, while continuously-labeled immunofluorescent probes can be…
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
Are the controls in AMPIPROBE® NG/CT/TV Extraction Controls, ENZ-GEN208-0500 quantitative or qualitative?
AMPIPROBE® NG/CT/TV Extraction Controls (ENZ-GEN208) are only qualitative and designed te used during genomic DNA extraction and is the qPCR…
Tue, Jul 18, 2023
What is the recommended thickness of the section to perform ISH?
Ideal section thickness depends on fixation and embedding method, as well as microscope that is available. FFPE tissue is ideally…
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