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Related Concept Videos

Transducer Mechanism: Nuclear Receptors01:31

Transducer Mechanism: Nuclear Receptors

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Nuclear receptors, or NRs, are unique transcription factors that regulate gene transcription and affect the cellular pathways involved in reproduction, development, or metabolism. Their ability to be stimulated by small lipophilic ligands and control vital cellular processes makes them ideal drug targets. Nearly 10-15% of currently prescribed drugs target these receptors.
About 48 different soluble family members of nuclear receptors are identified that can be divided into two main classes:
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Nuclear Export of mRNA02:31

Nuclear Export of mRNA

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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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Nuclear Stability03:18

Nuclear Stability

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Protons and neutrons, collectively called nucleons, are packed together tightly in a nucleus. With a radius of about 10−15 meters, a nucleus is quite small compared to the radius of the entire atom, which is about 10−10 meters. Nuclei are extremely dense compared to bulk matter, averaging 1.8 × 1014 grams per cubic centimeter. If the earth’s density were equal to the average nuclear density, the earth’s radius would be only about 200 meters.
To hold positively charged protons together...
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Reliability and Validity01:29

Reliability and Validity

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Reliability and validity are two important considerations that must be made with any type of data collection. Reliability refers to the ability to consistently produce a given result. In the context of psychological research, this would mean that any instruments or tools used to collect data do so in consistent, reproducible ways.
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Nuclear Fusion02:45

Nuclear Fusion

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The process of converting very light nuclei into heavier nuclei is also accompanied by the conversion of mass into large amounts of energy, a process called fusion. The principal source of energy in the sun is a net fusion reaction in which four hydrogen nuclei fuse and ultimately produce one helium nucleus and two positrons.
A helium nucleus has a mass that is 0.7% less than that of four hydrogen nuclei; this lost mass is converted into energy during the fusion. This reaction produces about...
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Non-nuclear Inheritance01:29

Non-nuclear Inheritance

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Most DNA resides in the nucleus of a cell. However, some organelles in the cell cytoplasm⁠—such as chloroplasts and mitochondria⁠—also have their own DNA. These organelles replicate their DNA independently of the nuclear DNA of the cell in which they reside. Non-nuclear inheritance describes the inheritance of genes from structures other than the nucleus.
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Related Experiment Video

Updated: Jan 25, 2026

Anti-Nuclear Antibody Screening Using HEp-2 Cells
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Antibody Validation Strategy for Nuclear Receptors.

Borbala Katona1, Ahmed Ibrahim2, Mårten Sundberg3

  • 1Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.

Methods in Molecular Biology (Clifton, N.J.)
|May 2, 2019
PubMed
Summary

Thorough antibody validation is crucial for reliable nuclear receptor research. This study presents a robust method using tissue microarrays and multiple techniques to ensure antibody specificity and performance, preventing flawed interpretations.

Keywords:
AntibodyEstrogen receptorImmunohistochemistryImmunoprecipitationMass spectrometryNuclear receptorsRNA-SeqTissue microarrayValidationWestern blotting

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High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum
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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Antibodies are essential for detecting nuclear receptors but suffer from cross-reactivity and batch-to-batch variability.
  • Inconsistent antibody validation leads to unreliable results and reproducibility issues in biological assays.

Purpose of the Study:

  • To present a comprehensive antibody validation scheme specifically designed for nuclear receptors.
  • To address the critical need for reliable antibody performance in research.

Main Methods:

  • Utilizing a validation tissue microarray (TMA) with characterized positive and negative controls.
  • Correlating immunohistochemical (IHC) staining with mRNA levels across diverse tissues.
  • Employing Western blotting (WB), immunoprecipitation (IP), and mass spectrometry (MS) to assess target binding specificity.

Main Results:

  • The proposed method enables rigorous assessment of antibody performance across various applications and sample types.
  • Validation through correlation of IHC and mRNA data provides a robust measure of antibody accuracy.
  • Specificity is confirmed by evaluating binding to intended and non-intended targets via WB, IP, and MS.

Conclusions:

  • A systematic approach to antibody validation is essential for accurate nuclear receptor detection.
  • The described validation scheme enhances the reliability and reproducibility of antibody-based assays.
  • Implementing this method ensures the integrity of research findings dependent on antibody specificity.