Related Experiment Video
Updated: Jan 30, 2026
09:30
Synthetic Antigen Controls for Immunohistochemistry
Published on: August 23, 2021
3.0K
Selecting an Optimal Positive IHC Control for Verifying Antigen Retrieval
Seshi R Sompuram1, Kodela Vani1, Anika K Schaedle1
1Medical Discovery Partners LLC, Boston, Massachusetts.
Summary
Selecting appropriate positive controls is crucial for verifying heat-induced epitope retrieval (HIER) in immunohistochemistry (IHC). Low or intermediate analyte concentration controls are best for confirming HIER efficacy, unlike high concentrations which can be insensitive.
Area of Science:
- * Immunohistochemistry
- * Molecular Biology
- * Pathology
Background:
- * Positive controls in immunohistochemistry (IHC) aim to detect issues in immunostaining and heat-induced epitope retrieval (HIER).
- * The specific features of a control essential for verifying HIER remain unclear.
- * Formalin-fixed tissues may not always be suitable for verifying HIER, as some stain without it.
Purpose of the Study:
- * To investigate the critical features of positive controls for verifying HIER in IHC.
- * To understand the impact of formalin fixation on epitope masking and HIER verification.
- * To determine the optimal analyte concentration in positive controls for effective HIER assessment.
Main Methods:
- * Utilized formalin-fixed peptide epitopes as a model system for precise control definition.
- * Analyzed variable epitope masking caused by formalin fixation based on primary antibody recognition.
- * Investigated the role of amino acid reactivity with formaldehyde in epitope masking variability.
Main Results:
- * Formalin fixation causes variable epitope masking, dependent on the specific epitope and primary antibody.
- * Amino acid composition influences an epitope's susceptibility to formaldehyde-induced masking.
- * Positive controls with low or intermediate analyte concentrations are more sensitive for verifying HIER than high concentrations.
Conclusions:
- * Not all formalin-fixed tissues are adequate controls for HIER verification.
- * Epitope masking by formalin fixation varies significantly.
- * Selecting positive controls with appropriate analyte concentration is critical for validating HIER in IHC.
Related Concept Videos
01:12Retrieval
444
Retrieval is the process of getting information out of memory storage and back into conscious awareness. This ability is essential for daily tasks like brushing hair and teeth, driving to work, and performing job duties. Retrieval occurs in three ways: recall, recognition, and relearning.
Recall involves accessing information without cues, such as during an essay test, where individuals must retrieve facts and concepts from memory unaided. Another example is remembering the name of a colleague...
Recall involves accessing information without cues, such as during an essay test, where individuals must retrieve facts and concepts from memory unaided. Another example is remembering the name of a colleague...
444
01:45ER Retrieval Pathway
4.8K
In the secretory pathway, vesicles transport proteins from one cellular compartment to another in forward transport to deliver the protein to its correct location. Occasionally, misfolded proteins and incorrect proteins escape their original compartments, and a retrieval pathway is used to return the escaped proteins to their original compartment.
The ER uses many checkpoints to prevent the entry of incorrectly folded or a resident protein as cargo onto a transport vesicle. These mechanisms...
The ER uses many checkpoints to prevent the entry of incorrectly folded or a resident protein as cargo onto a transport vesicle. These mechanisms...
4.8K
02:32Position-effect Variegation
7.1K
In 1928, a German botanist Emil Heitz observed the moss nuclei with a DNA binding dye. He observed that while some chromatin regions decondense and spread out in the interphase nucleus, others do not. He termed them euchromatin and heterochromatin, respectively. He proposed that the heterochromatin regions reflect a functionally inactive state of the genome. It was later confirmed that heterochromatin is transcriptionally repressed, and euchromatin is transcriptionally active chromatin.
7.1K
01:32What is Natural Selection?
128.9K
Natural selection is an evolutionary process in which individuals with survival-promoting traits reproduce at higher rates. These favorable traits become more common within a population or species. Naturally selected traits initially arise via random genetic mutations. In order for selection to occur, there must be variation within a population, the trait controlling the variation must be heritable, and there must be an evolutionary advantage for variation in the trait.
128.9K
00:48Optimal Foraging
13.8K
How animals obtain and eat their food is called foraging behavior. Foraging can include searching for plants and hunting for prey and depends on the species and environment.
13.8K
01:03Serial Position Effect
542
The serial position effect is a cognitive phenomenon where individuals are more likely to recall the first and last items in a list compared to those in the middle. This effect is divided into the primacy effect and the recency effect. The primacy effect is observed when the initial items in a list are remembered better. This occurs because these items are rehearsed more frequently or receive more elaborative processing, allowing them to be encoded into long-term memory more effectively. For...
542