Antibody-based Quantification
Antibody-based Quantification - Related Videos
05:55
Quantification of Autoreactive Antibodies in Mice upon Experimental Autoimmune Encephalomyelitis
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Cited by 3 •
2023
Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS), which shares with the human disease a robust humoral autoimmune response. Here, we report a simple and flexible ELISA protocol to quantify autoantibodies in the serum of EAE immunized mice.
04:43
Antibody Conjugated Nanoparticle-based Quantification Using Dark Field Microscopy: A Procedure to Capture and Quantify Specific Exosomes from Body Fluids
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2023
This video describes the use of low-magnification, dark-field microscopy to quantify specific exosomes in small volume biofluidic samples. The exosomes thus identified can serve as potential cancer biomarkers.
07:26
Virus Propagation and Cell-Based Colorimetric Quantification
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2023
The present protocol describes the propagation of Zika virus (ZIKV) in Vero African green monkey kidney cells and the quantification of ZIKV using cell-based colorimetric immunodetection methods in 24-well and 96-well (high throughput) formats.
04:24
An Open-Source Framework for Mass Calculation of Antibody-Based Therapeutic Molecules
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Cited by 1 •
2023
This article describes the use of a software application, mAbScale, for the calculation of masses for monoclonal antibody-based protein therapeutics.
10:23
Standardization of a Cytometric Bead Assay Based on Egg-Yolk Antibodies
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Cited by 1 •
2023
This protocol describes a methodology for the preparation of latex beads for assays using IgY antibodies for antigen detection.
FAQs
Selectivity comes from the antibody’s ability to recognize the intended molecule within a sample. This narrows the measurement toward the target rather than treating all sample constituents as equivalent. In biochemical analysis, this focused recognition can support measurement of proteins, hormones, drugs, or other antigens, including when direct measurement is difficult.
Known-concentration standards provide the reference needed to interpret an assay signal quantitatively. The unknown sample’s response is compared with these reference values, allowing its target abundance to be determined rather than reported only as detectable or undetectable. This calibration step also supports comparisons among samples analyzed with the same measurement approach.
An enzyme, fluorescent label, or other detectable marker converts the antibody-target interaction into a measurable signal. The choice described in the assay determines how the binding event becomes observable, but quantitative interpretation still depends on comparing that signal with known standards. Thus, labeling supplies detectability, whereas antibody recognition supplies target selectivity.
Specificity and sensitivity contribute different strengths to an antibody-based measurement. Specificity helps focus the assay on the intended antigen, while sensitivity supports detection when direct measurement is difficult. Their combination is especially useful in complex biochemical samples, where the target must be distinguished and compared across preparations.
A basic workflow begins with a sample containing the molecule of interest and an antibody selected for recognition. The target binds the antibody, a detectable marker generates the assay signal, and standards of known concentration provide the comparison scale. The measured sample response is then interpreted against those standards to estimate target abundance.
Applications span protein characterization, diagnostics, pharmacology, and cell biology. In biochemistry, the same measurement logic can be applied to proteins, hormones, drugs, and other antigens, depending on the antibody and assay design. Results can reveal target abundance or enable comparisons between samples, making the approach useful across research and applied settings.