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Can BIO-PROBE®, PATHO-GENE®, or AMPIVIEW® ready-to-use ISH probes be used for FISH?
Biotin-labeled in situ probes are most commonly used for chromogenic detection, they can easily be adapted to fluorescent detection with…
Tue, Jul 18, 2023
How important are denaturation and hybridization temperature in ISH?
Correct denaturation and hybridization temperatures are paramount for successful ISH. Regardless of the probe or target being RNA or DNA,…
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
Why are peptidases/proteinases used in ISH?
Peptidases or proteinases like proteinase K, pepsin or trypsin are used in ISH to digest proteins contained in the sample…
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
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
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 ISH probes typically labeled with?
Common labels include biotin, digoxigenin, and fluorochromes. Biotin is detected with streptavidin- conjugated enzymes followed by an enzymatic colorimetric reaction.…
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