Hit Confirmation
Hit Confirmation - Related Videos
01:31
Confirmation Biases
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2020
The confirmation bias is the tendency to focus on information that confirms our existing beliefs and ignore information that is inconsistent with our expectations. For example, if you think that your professor is not very nice, you notice all of the instances of rude behavior exhibited by the professor while ignoring the countless pleasant interactions he is involved in on a daily basis. Have you ever fallen prey to the confirmation bias, either as the source or target of such bias? Another...
06:19
High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography
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Cited by 16 •
2023
This protocol details high-throughput crystallization screening, ranging from the 1,536 microassay plate preparation to the end of a 6 week experimental time window. Details are included about the sample setup, the imaging obtained, and how users can perform analyses using an artificial intelligence-enabled graphical user interface to quickly and efficiently identify macromolecular crystallization conditions.
01:34
Humane Euthanasia and Confirmation of Death in Rodents
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2026
All animal procedures described here must be conducted in accordance with institutional animal ethics guidelines and approved by IACUC. All procedures must follow the principles of the 3Rs—Replacement, Reduction, and Refinement—and must be performed by trained personnel. Humane endpoints in rodents are predefined clinical, behavioral, or physiological indicators that signal an animal is experiencing significant pain, distress, or declining health. When these signs are observed, timely...
02:27
Confirming Viral Transduction in Recellularized Rat Liver Using Immunohistochemical Staining
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2026
Source: Hiller, T. et. al. Study of Viral Vectors in a Three-dimensional Liver Model Repopulated with the Human Hepatocellular Carcinoma Cell Line HepG2. J. Vis. Exp. (2016)This video demonstrates immunohistochemical staining to evaluate viral gene transfer in a recellularized rat liver model. Reporter expression, confirmed by antibody detection and co-localization with nuclear staining, indicates successful viral transduction in the cells.
10:41
Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
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Cited by 2 •
2017
A construct encoding TMEM184A with a GFP tag at the carboxy-terminus designed for eukaryotic expression, was employed in assays designed to confirm the identification of TMEM184A as a heparin receptor in vascular cells.
FAQs
Replicate testing shows whether a candidate produces a reproducible signal rather than an isolated result. Measuring responses across concentrations adds evidence about how activity changes with dose, helping researchers judge the reliability of the hit and identify candidates whose behavior merits further investigation in subsequent studies.
Retesting alone may not resolve why a signal appears. Orthogonal assays and counter-screens provide additional checks for whether observed activity reflects the intended biological mechanism, rather than assay interference or nonspecific effects. This broader evaluation makes the evidence more credible before a compound is treated as a meaningful research lead.
Toxicity can make an apparent response difficult to interpret as mechanism-specific activity. During confirmation, researchers therefore consider whether the signal persists as a biologically meaningful effect rather than reflecting compound toxicity or a broad nonspecific action. This distinction helps prioritize compounds with more credible and selective activity for later work.
A practical sequence begins by retesting each candidate in replicate, followed by measurements across concentrations. Researchers then apply orthogonal assays or counter-screens to examine the source and specificity of the response. Results from these checks determine whether the candidate can advance as a credible lead for target validation or structure–activity studies.
It is used after screening has produced candidate hits and before compounds move into hit-to-lead optimization. The stage helps researchers prioritize candidates that show reproducible, concentration-dependent, selective, and biologically meaningful activity. By distinguishing misleading signals early, confirmation supports more efficient downstream research and better-informed advancement decisions.
Confirmed activity can support target validation by showing that a compound’s response is sufficiently credible to investigate in relation to a biological target. It also provides a foundation for structure–activity studies and hit-to-lead optimization. In this way, confirmation connects an initial screening result with decisions about which compounds merit deeper biological investigation and development.