Caco 2 cell permeability assay
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Creative Bioarray is offering Caco-2 permeability assay to help determine the absorption and the bioavailability of drug candidates, facilitating the lead optimization process in drug discovery. https://www.creative-bioarray.com/Services/caco-2-permeability-assay.htm
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Comparison of caco 2 with other cell-based models for intestinal permeability...
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This document summarizes a seminar on using Caco-2 monolayers to study transport across the intestinal barrier. The Caco-2 cell line spontaneously differentiates into enterocytes that form a polarized monolayer mimicking intestinal absorption. Key aspects covered include Caco-2 cell culture and characterization, permeability assays to measure transport, and applications in drug development. Validation is done using reference compounds to classify drug absorption. Considerations for biological factors and analytical methods are also discussed.
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Caco-2 cell permeability assay is the golden standard for in vitro prediction of intestinal drug permeability and absorption.
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OPG expression in intestinal epithelial cells appears to be suppressed when exposed to commensal bacteria. OPG binds to both live and killed bacteria and this bound OPG can be released through detergent treatment. Treatment of Caco 2 cell supernatants with detergent does not increase detectable OPG levels by ELISA, indicating ELISA detects the true OPG concentration.
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Adult T. confusum beetles were collected using various methods and preserved under different conditions to test the quality of extracted DNA for barcoding. Beetles preserved in 100% ethanol at -20°C produced high quality DNA, while those in water at room temperature produced degraded DNA. Preservation in 70% ethanol or dry collection with dichlorvos strips had minimal negative effects on DNA compared to 100% ethanol or wet collection. Low temperature storage is important for maintaining DNA quality for barcoding.
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MuSeek is a MuA transposon-based library preparation
method that offers a streamlined workflow and the sensitivity
necessary for low input experiments. We are exploring
improvements that reduce required processing steps and
enhance performance.
We present data using cell lysates as direct input into the
library preparation reaction:
• A single bacterial colony for whole genome sequencing
(WGS)
• Single human cells for low-pass aneuploidy detection
These improvements to MuSeek offer exciting new
possibilities for simple, rapid, and low input library preparation
up front of the Ion Torrent™ Sequencing Systems.micronucleus assay.pptx
This document presents a procedure for conducting genotoxicity studies using in vitro and in vivo micronucleus assays. It introduces the micronucleus test as a method used in toxicological screening to identify potential genotoxic compounds. The procedure involves growing mammalian cells on cover glasses, treating the cells with test compounds, fixing and staining the cells, and examining them under a fluorescence microscope to detect micronuclei formation indicating DNA damage. References are provided for additional information on micronucleus testing methodology and applications.
Ex Bio 2016 poster
1) The document evaluates sample disruption techniques for extracting live E. coli and recombinant DNA from food using a bead mill and rotor stator homogenizer. Both techniques recovered similar amounts of viable cells as controls and allowed detection of E. coli down to 1x10-6 CFU/ml by PCR.
2) Spinach and beef samples inoculated with known levels of fluorescent E. coli were processed with each homogenizer. Live cell recovery was evaluated by plating and PCR detection limits were analyzed. A linear relationship was observed between CFU levels and DNA amounts amplified by PCR.
3) Both homogenizers fully disrupted samples without compromising bacterial viability, recovering up to 82% of cells. This vigorous processing could
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manish.pptx
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Carbohydrate, isolation and purification techniques. A complete view.
Lab scale step by step isolation, purification and optimization of bio-molecules with complete details of techniques and equipment used.
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This document describes a procedure for isolating genomic DNA from bacterial cells. It involves growing a culture of bacterial cells, harvesting and lysing the cells to release the DNA. The DNA is then purified using phenol-chloroform extraction and precipitated with alcohol. The extracted DNA is analyzed on an agarose gel and quantified. Alternative methods are also discussed, including using DNAzol or sonication for cell lysis and silver nanoparticles or TE buffer for DNA extraction and purification. The overall procedure allows for the isolation of pure bacterial genomic DNA for use in downstream molecular biology applications.
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Mammalian project poster
Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions. Mammalian cell culture technology has become a major field in modern biotechnology; mammalian cell culture refers to the cells of a mammalian, isolated from specific tissues (i.e. skin, liver, glands, etc.) and further cultivated and reproduced in an artificial medium. Cell culture technology is currently playing a major role in toxicity testing, cancer research, virology, genetic engineering, and gene therapy.
OBJECTIVE:
To observe the transfection of CHO and HEK cells with GFP
To observe the recombinant GFP using Western Blotting
To purify the transfected HEK and CHO cells using AKTA Pure Purification
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2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
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Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
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Caco 2 cell permeability assay
- 2. Contents Caco-2 Monolayer Caco-2 Cell Culture Measuring TEER Assay Procedure Papp Calculation
- 3. 3Part 1 1.1 Characteristics of Caco-2 Cells 1 Origin: Human colorectal adenocarcinoma 2 Growth in culture: Monolayer epithelial cells 3 Differentiation: 21 days after confluence in standard culture medium 4 Morphology: Polarised cells, with tight junctions, apical, brush border
- 4. 4Part 1 1.2 Advantages of Caco-2 Monolayer 01 Spontaneously differentiate to express morphological and functional characteristics of mature small-intestinal enterocytes 02 Four times higher in trans-epithelial resistance compared to HT 29-cell monolayer 03 Expressing various drug metabolizing enzymes such as aminopeptidase, esterase, and sulfatase.
- 5. 5Part 1 1.3 Limitation of Caco-2 Monolayer Limitation of Caco-2 P-gp LC/MS Time Cost Enzyme Mucus Tissue
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- 8. 8Part 1 3.1 Measuring TEER 01 Washing the cell monolayer with 37℃ tempered D- PBS (with Ca2+, Mg2+) Transport media was added into the apical and the basal chamber. Allow for equilibrium for 60 min in the cell culture incubator. After measurement, Caco-2 monolayer with TEER values exceeding 250 Ω x cm2 were used for transport experiments. Background TEER may be recorded in wells without cell monolayers, and can be subtracted from the raw TEER values with cells. 02 03 04
- 9. 9Part 1 4.1 Caco-2 Permeability Assay Procedure A B C D Wash Wash the monolayer with sterile HBSS, pH7.4, and transfer the filter plate to a 12- well transport analysis plate. Add analytes Add the test compounds to the filter well. Drug concentrations typically ranging from 10 μm to 200 μm. Incubate Join the filter and receiver plates once all drugs and buffer have been added. Incubate at 37℃ shaking at 60 rpm on a rotary shaker. For analysis At the end of the incubation, remove a fixed volume (typically 50-100 μL) directly from the apical and basolateral wells to a clean plate.
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