Supernatant
Supernatant - Related Videos
01:49
An Assay To Evaluate the Anticancer Effects of Lactobacillus Cell-Free Supernatant
0 Views •
2025
This video demonstrates an in vitro assay to study the anticancer effects of Lactobacillus cell-free supernatant (LCFS). Upon exposing colorectal cancer spheroids to varying doses of LCFS, the interaction with the bacterial metabolites in LCFS induces apoptosis in the cancer cells, disrupting the spheroid structure and leading to cell death in a dose-dependent manner.
03:40
Heparin Affinity Chromatography-Based Baculovirus Purification: A Technique to Isolate Baculovirus From Insect Cell Supernatant
0 Views •
2025
In this video, we demonstrate the technique to isolate baculovirus from an insect cell supernatant using heparin affinity chromatography. The isolation is facilitated by the affinity between the heparin in the column to the baculovirus envelope glycoprotein.
06:07
Evaluating Cell Death Using Cell-Free Supernatant of Probiotics in Three-Dimensional Spheroid Cultures of Colorectal Cancer Cells
0 Views •
Cited by 15 •
2020
Here methods are presented to understand anti-cancer effects of Lactobacillus cell-free supernatant (LCFS). Colorectal cancer cell lines show cell deaths when treated with LCFS in 3D cultures. The process of generating spheroids can be optimized depending on the scaffold and the analysis methods presented are useful for evaluating the involved signaling pathways.
08:36
Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
0 Views •
Cited by 7 •
2018
Real-time quantitative polymerase chain reaction analysis combined with reverse transcription (RT-qPCR) has been widely used to measure the level of RNA virus infections. Here we present a direct RT-qPCR assay, which does not require an RNA purification step, developed for the quantification of several RNA viruses, including dengue virus.
03:14
Obtaining Cytotoxic Amyloids Following Bacterial Infection of Rat Pulmonary Endothelial Cells
0 Views •
2025
Source: Balczon, R., et al. Methods for Detecting Cytotoxic Amyloids Following Infection of Pulmonary Endothelial Cells by Pseudomonas aeruginosa. J. Vis. Exp. (2018)This video demonstrates the procedure for generating cytotoxic supernatant from pulmonary endothelial cells following infection with Pseudomonas aeruginosa. It outlines how bacterial effector proteins disrupt host cell structure and induce the release of cytotoxic amyloid oligomers into the supernatant.
FAQs
During centrifugation, increased centrifugal force promotes movement of cells, organelles, precipitates, or other dense particles toward the bottom. Soluble components remain in the liquid above that material, creating a fraction enriched for dissolved molecules rather than sedimented structures. This physical separation connects sample preparation with downstream measurement.
Particles that are not fully separated can remain in the liquid fraction and introduce particulate contamination into an assay. Careful separation helps produce a more clarified sample, allowing measurements to reflect dissolved proteins, metabolites, nucleic acids, enzymes, or other molecules more consistently. The quality of this step therefore supports reproducible analysis.
The pellet contains material driven to the bottom during separation, including cells, organelles, precipitates, or other dense components. The Supernatant is used when the experimental question concerns soluble or dissolved contents that remain above that material. Comparing these fractions can therefore distinguish molecular information associated with the liquid phase from sedimented material.
A sample is first subjected to a separation step, commonly after cell culture, cell lysis, or biochemical extraction. Once dense material has settled or formed a pellet, the liquid fraction above it is transferred for analysis. Careful handling during transfer is important because disturbing the sediment can increase particulate contamination and reduce consistency.
Researchers can use this fraction in assays measuring secreted proteins, metabolites, nucleic acids, enzymes, or other dissolved molecules. The appropriate target depends on whether the original sample came from cell culture, lysis, or biochemical extraction. Results can provide information about molecular composition or cellular activity represented in the liquid phase.
The liquid fraction is especially useful when researchers need to examine materials released by cells or molecules extracted into a solution without analyzing the separated sediment. In cell culture, it can support measurement of secreted components; after lysis or biochemical extraction, it can be analyzed for dissolved molecular contents. This makes it valuable for studying cellular activity and sample composition.