Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Methods of reducing fever01:22

Methods of reducing fever

1.4K
The signs and symptoms of fever include hot and dry skin, flushed face, thirst, muscle aches, anorexia, headache, tachycardia, tachypnea, and fatigue. Elevated body temperature is reduced using two methods: pharmacological and nonpharmacological. Proper identification and treatment of the root cause of a fever is of utmost importance.
Pharmacological Methods of Reducing Fever:
1.4K
Background and Environment Affect Phenotype02:27

Background and Environment Affect Phenotype

7.7K
Although the genetic makeup of an organism plays a major role in determining the phenotype, there are also several environmental factors, such as temperature, oxygen availability, presence of mutagens, that can alter an organism’s phenotype.
An example of how genetic background affects phenotype can be seen in horses. The Extension gene in horses is responsible for their coat color. A wild-type gene (EE) produces black pigment in the coat, while a mutant gene (ee) produces red pigment. A...
7.7K
Difference from Background: Limit of Detection01:05

Difference from Background: Limit of Detection

8.4K
The limit of detection (LOD) is the smallest amount of analyte that can be distinguished from the background noise. The LOD value corresponds to the concentration at which the analyte signal is three times larger than the standard deviation of the blank signal. Below this value, the analyte signal cannot be differentiated from the background noise. It is calculated by dividing the calibration slope by 3 times the standard deviation of the blank signals.
The LOD indicates the presence or absence...
8.4K
Reducing Line Loss01:18

Reducing Line Loss

395
In a three-phase circuit, line loss is an indicator of energy dissipated as heat due to the resistance of transmission lines. To address this, incorporating transformers into the system—a step-up transformer at the source and a step-down transformer at the load—is a strategic solution. Two three-phase transformers are introduced to improve this.
With a step-up transformer at the source, the voltage is increased, thereby reducing the current in the transmission lines since power loss in...
395
Adaptations that Reduce Water Loss01:57

Adaptations that Reduce Water Loss

28.3K
Though evaporation from plant leaves drives transpiration, it also results in loss of water. Because water is critical for photosynthetic reactions and other cellular processes, evolutionary pressures on plants in different environments have driven the acquisition of adaptations that reduce water loss.
28.3K
Role of Reduced Coenzymes NADH and FADH₂01:29

Role of Reduced Coenzymes NADH and FADH₂

17.1K
The energy released from the breakdown of the chemical bonds within nutrients can be stored either through the reduction of electron carriers or in the bonds of adenosine triphosphate (ATP). In living systems, a small class of compounds functions as mobile electron carriers, molecules that bind to and shuttle high-energy electrons between compounds in pathways. The principal electron carriers that will be considered originate from the B vitamin group and are derivatives of nucleotides; they are...
17.1K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Metabolic state modulates risky foraging behavior via multiple branches of the insulin/IGF-1-like pathway in C. elegans.

G3 (Bethesda, Md.)·2026
Same author

Characterization of deglycosylated anti-cocaine mAb by differential scanning fluorimetry and fluorescence labeling.

bioRxiv : the preprint server for biology·2025
Same author

Assessment of Lectin Staining Biomarkers using a Murine Model of GNE Myopathy.

microPublication biology·2025
Same author

Synthesis and characterization of a functional benzoylecgonine - bovine serum albumin conjugate for quantification of a humanized monoclonal anti-cocaine antibody.

Biochemistry and biophysics reports·2025
Same author

Selective real-time disulfide reduction with simultaneous fluorescent labeling of a humanized anti-cocaine mAb.

Biochemistry and biophysics reports·2025
Same author

Creatine Exposure in <i>Caenorhabditis elegans</i> Induces Physiological and Morphological Changes without Metabolic Conversion to Creatinine: A Novel HPLC Analysis.

ACS omega·2025
Same journal

Optimized intracellular flow cytometry panel enables CD4 and CD8 T cell cytokine profiling in Syrian hamsters.

Journal of immunological methods·2026
Same journal

Isosulfan blue sentinel lymph node biopsy enables reliable lymph node harvest and multicolor flow cytometry in mice.

Journal of immunological methods·2026
Same journal

Type-specific antibody detection of herpes simplex virus types 1&2 (HSV-1&2) in fingerstick blood at point-of-care sites by a rapid and sensitive lateral flow immunochromatographic assay.

Journal of immunological methods·2026
Same journal

Development of chimeric DGP-IgG antibodies as quality control for celiac disease diagnosis.

Journal of immunological methods·2026
Same journal

Stepwise single-cell-resolved deep immunophenotyping pipeline to characterise immune heterogeneity and functionality in health and disease.

Journal of immunological methods·2026
Same journal

Performance evaluation of Meso Scale Discovery (MSD) quantitative serological assays for detection of binding (IgG, IgA, IgM) and ACE2 inhibitory antibody levels for SARS-CoV-2.

Journal of immunological methods·2026
See all related articles

Related Experiment Video

Updated: Feb 14, 2026

Computer-based Multitaper Spectrogram Program for Electroencephalographic Data
04:13

Computer-based Multitaper Spectrogram Program for Electroencephalographic Data

Published on: November 13, 2019

12.9K

Evaluation of methods to reduce background using the Python-based ELISA_QC program.

Rose P Webster1, Cinder F Cohen1, Fatima O Saeed1

  • 1Department of Pharmacology and Systems Physiology, University of Cincinnati, College of Medicine, 231 Albert Sabin Way, Cincinnati, OH 45267-0575, United States.

Journal of Immunological Methods
|February 23, 2018
PubMed
Summary
This summary is machine-generated.

A novel immunodepletion (ID) method and heparin (HP) effectively reduce high backgrounds in enzyme-linked immunosorbent assay (ELISA) using biotinylated antibodies. The ID method offers superior results for quantitative protein analysis in immunological assays.

Keywords:
Anti-cocaine antibodyBackgroundELISAHeparin washImmunodepletionLevey-Jennings chartNon-specific reactivity

More Related Videos

Applying an eMASS Customization Program as a Research Tool to Evaluate Consumer Benefits
08:27

Applying an eMASS Customization Program as a Research Tool to Evaluate Consumer Benefits

Published on: September 27, 2019

7.3K
A Novel Digital Platform for a Monitored Home-based Cardiac Rehabilitation Program
04:24

A Novel Digital Platform for a Monitored Home-based Cardiac Rehabilitation Program

Published on: April 19, 2019

12.7K

Related Experiment Videos

Last Updated: Feb 14, 2026

Computer-based Multitaper Spectrogram Program for Electroencephalographic Data
04:13

Computer-based Multitaper Spectrogram Program for Electroencephalographic Data

Published on: November 13, 2019

12.9K
Applying an eMASS Customization Program as a Research Tool to Evaluate Consumer Benefits
08:27

Applying an eMASS Customization Program as a Research Tool to Evaluate Consumer Benefits

Published on: September 27, 2019

7.3K
A Novel Digital Platform for a Monitored Home-based Cardiac Rehabilitation Program
04:24

A Novel Digital Platform for a Monitored Home-based Cardiac Rehabilitation Program

Published on: April 19, 2019

12.7K

Area of Science:

  • Immunology
  • Biotechnology
  • Assay Development

Background:

  • High background due to non-specific reactivity is a common challenge in quantitative immunological assays like ELISA, immunohistochemistry, and Western blot.
  • Biotinylated antibodies are frequently used in these assays, but certain lots can exhibit elevated background noise, impacting data accuracy.
  • Optimizing assay conditions and exploring novel blocking strategies are crucial for reliable protein quantification.

Purpose of the Study:

  • To quantitatively compare different methods for reducing background noise in enzyme-linked immunosorbent assay (ELISA) using commercial biotinylated antibodies.
  • To evaluate the efficacy of heparin (HP) and a novel immunodepletion (ID) method in mitigating non-specific reactivity.
  • To assess the performance of these methods using a recombinant humanized anti-cocaine monoclonal antibody and the Python-based ELISA_QC program.

Main Methods:

  • Systematic comparison of traditional background reduction techniques (e.g., adjusting incubation time, blocking agent concentration, secondary antibody dilution) with heparin (HP) and immunodepletion (ID) methods.
  • Application of a recombinant humanized anti-cocaine monoclonal antibody in an ELISA format.
  • Quantitative analysis of background reduction and assay performance using the Python-based ELISA_QC program.

Main Results:

  • Both heparin (HP) at 1 μg/ml and the immunodepletion (ID) method significantly reduced elevated background absorbance values in ELISA.
  • HP reduced background from a mean of 0.313 ± 0.015 to 0.137 ± 0.002, while ID reduced it from 0.331 ± 0.010 to 0.146 ± 0.013.
  • The ID method demonstrated more consistent results across the ELISA standard curve compared to the HP method, as analyzed by ELISA_QC.

Conclusions:

  • The immunodepletion (ID) method provides a superior solution for resolving high background issues associated with specific lots of biotinylated secondary antibodies in ELISA.
  • While more labor-intensive, the ID method yields more reliable and reproducible quantitative data in immunological assays.
  • These findings offer valuable insights for optimizing ELISA protocols and improving the accuracy of protein quantification in research and diagnostics.