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Our application notes demonstrate the quantitation of nucleic acids and protein in a microplate format offers higher throughput and automated calculation of results compared to other methods.
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DNA/RNA Quantitation
The absorbance of a DNA sample measured at 260 nm on a spectrophotometer or microplate reader can be used to calculate its concentration. Absorbance quantitation works on samples ranging from about 0.25 ug/mL to about 125 ug/mL in a microplate format. Some instrumentation enables the quantitation of very small sample volumes, as little as 2 uL. When greater sensitivity is required, fluorescence methods allow quantitation of as little as a few picograms of DNA.
DNA/RNA Quantitation
The absorbance of a DNA sample measured at 260 nm on a spectrophotometer or microplate reader can be used to calculate its concentration. Absorbance quantitation works on samples ranging from about 0.25 ug/mL to about 125 ug/mL in a microplate format. Some instrumentation enables the quantitation of very small sample volumes, as little as 2 uL. When greater sensitivity is required, fluorescence methods allow quantitation of as little as a few picograms of DNA.
ELISA
Enzyme-linked immunosorbent assays (ELISAs) are used to measure the amount of a specific protein, typically in a microplate format, and results are most often detected via absorbance in the visible wavelength range. Chemiluminescent and fluorescent ELISA formats offer enhanced sensitivity for accurate quantitation of less abundant analytes.
ELISA
Enzyme-linked immunosorbent assays (ELISAs) are used to measure the amount of a specific protein, typically in a microplate format, and results are most often detected via absorbance in the visible wavelength range. Chemiluminescent and fluorescent ELISA formats offer enhanced sensitivity for accurate quantitation of less abundant analytes.
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Enzyme Activity
Enzyme activity can be monitored in real time using a chromogenic substrate which, upon its addition to the enzyme, produces a change in color that is detectable on an absorbance microplate reader. Using an instrument with onboard liquid handling capability, substrate can be added automatically while the reaction is monitored, enabling detection of the earliest part of the reaction and thus more accurate estimates of binding affinity.
Enzyme Activity
Enzyme activity can be monitored in real time using a chromogenic substrate which, upon its addition to the enzyme, produces a change in color that is detectable on an absorbance microplate reader. Using an instrument with onboard liquid handling capability, substrate can be added automatically while the reaction is monitored, enabling detection of the earliest part of the reaction and thus more accurate estimates of binding affinity.
Fluorescent Protein Detection
Fluorescent proteins have become enormously popular as tools for monitoring biological events in vivo. In addition to green fluorescent protein (GFP) from the jellyfish Aequorea victoria, there are now numerous others available from other species of jellyfish and reef coral. These proteins can be expressed in a diverse range of cells and organisms, where they are used to track many cellular processes, including protein synthesis and translocation, gene induction, and cell lineage.
Fluorescent Protein Detection
Fluorescent proteins have become enormously popular as tools for monitoring biological events in vivo. In addition to green fluorescent protein (GFP) from the jellyfish Aequorea victoria, there are now numerous others available from other species of jellyfish and reef coral. These proteins can be expressed in a diverse range of cells and organisms, where they are used to track many cellular processes, including protein synthesis and translocation, gene induction, and cell lineage.
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Kinase/Phosphatase Assays
Kinases are one of the most critical targets in drug discovery today. These enzymes are key components in cellular signaling pathways, and disruption to their function causes a variety of diseases, such as metabolic diseases, certain cancers, and cardiac disease. The functionally related phosphatases and phosphodiesterases are also important screening targets.
Kinase/Phosphatase Assays
Kinases are one of the most critical targets in drug discovery today. These enzymes are key components in cellular signaling pathways, and disruption to their function causes a variety of diseases, such as metabolic diseases, certain cancers, and cardiac disease. The functionally related phosphatases and phosphodiesterases are also important screening targets.
Mycoplasma Monitoring
Mycoplasma, the smallest and simplest of the prokaryotes, are common contaminants of cell cultures. Symptoms of mycoplasma contamination include a reduction in the rate of proliferation and changes in cellular responses, including gene expression. Mycoplasma cannot be detected by simply examining cell cultures under a microscope, so a variety of methods such as fluorescence- and luminescence-based assays have been developed to enable researchers to monitor for contamination.
Mycoplasma Monitoring
Mycoplasma, the smallest and simplest of the prokaryotes, are common contaminants of cell cultures. Symptoms of mycoplasma contamination include a reduction in the rate of proliferation and changes in cellular responses, including gene expression. Mycoplasma cannot be detected by simply examining cell cultures under a microscope, so a variety of methods such as fluorescence- and luminescence-based assays have been developed to enable researchers to monitor for contamination.
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Protein Quantitation (Bradford, Lowry, BCA, DC)
Protein concentration can be measured directly, via absorbance at 280 nm in a UV spectrophotometer, or indirectly, using colorimetric methods such as BCA or Bradford assays. Absorbance quantitation is easy to do as it does not require additional reagents, but colorimetric methods offer greater sensitivity and are often preferred when samples are precious. Both can be performed in a UV-vis spectrophotometer or in an absorbance microplate reader.
Protein Quantitation (Bradford, Lowry, BCA, DC)
Protein concentration can be measured directly, via absorbance at 280 nm in a UV spectrophotometer, or indirectly, using colorimetric methods such as BCA or Bradford assays. Absorbance quantitation is easy to do as it does not require additional reagents, but colorimetric methods offer greater sensitivity and are often preferred when samples are precious. Both can be performed in a UV-vis spectrophotometer or in an absorbance microplate reader.
SNP Genotyping
Genotyping is a process for analyzing genetic differences among individuals by examining their DNA sequences. Single nucleotide polymorphisms (SNPs) are one of the most common types of genetic variation, consisting of a single nucleotide mutation at a specific locus. SNP genotyping has proven very useful in identifying disease-related mutations in various species, and as a result many techniques for SNP detection have been developed.
SNP Genotyping
Genotyping is a process for analyzing genetic differences among individuals by examining their DNA sequences. Single nucleotide polymorphisms (SNPs) are one of the most common types of genetic variation, consisting of a single nucleotide mutation at a specific locus. SNP genotyping has proven very useful in identifying disease-related mutations in various species, and as a result many techniques for SNP detection have been developed.
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Western Blot
Western blotting is a popular technique used for protein detection and quantitation. Learn about the various techniques used to detect proteins on western blot membranes including colorimetric, fluorescence and chemiluminescence. We’ll also introduce you to a novel, first-of-its-kind western blot detection system on a multi-mode microplate reader.
Western Blot
Western blotting is a popular technique used for protein detection and quantitation. Learn about the various techniques used to detect proteins on western blot membranes including colorimetric, fluorescence and chemiluminescence. We’ll also introduce you to a novel, first-of-its-kind western blot detection system on a multi-mode microplate reader.
Resources of Protein and Nucleic Acid Detection
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DNA and RNA absorbance measurements using SpectraMax Microplate Readers
DNA and RNA absorbance measurements using SpectraMax Microplate Readers
Ultraviolet (UV) measurements in microplates became possible when Molecular Devices introduced the first UV-capable microplate reader. Since then, microplate measurements of DNA, RNA, and…
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Using the NanoOrange Protein Kit with SpectraMax Microplate Readers
Using the NanoOrange Protein Kit with SpectraMax Microplate Readers
This application note describes how to use the NanoOrange® Protein Quantitation Kit from Life Technologies in SpectraMax® microplate readers with the fluorescence detection mode and SoftMax…
과학 포스터
Western Blot Protein Detection and Quantitation Based on Europium Labeled Proteins using a Plate Reader
Western Blot Protein Detection and Quantitation Based on Europium Labeled Proteins using a Plate Reader
Learn how the ScanLater Western Blot system can be used to quantify protein levels in samples using Europium-based antibodies on the…
eBook
Detect Nucleic Acids and Proteins Like a Pro
Detect Nucleic Acids and Proteins Like a Pro
Accurate and sensitive detection of nucleic acids and proteins are critical to many experiments.
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Protein quantitation with the EMax Plus Microplate Reader
Protein quantitation with the EMax Plus Microplate Reader
Endpoint readers are prolific in the laboratory since absorbance has become the detection of choice for many applications. Examples include ELISAs for quantitation of cytokines and protein…
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PathCheck applied to measurement of protein solutions in the SPECTRAmax® PLUS microplate spectrophotometer
PathCheck applied to measurement of protein solutions in the SPECTRAmax® PLUS microplate spectrophotometer
Protein concentration is commonly estimated by measuring the absorbance at 280
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ß-Lactamase-Inhibitor Protein Binding Affinities: Detection of Hydrolysis of ß-Lactam Substrate by TEM-1 Enzyme and Inhibitor Binding Using The FlexStation® 3 Microplate Reader
ß-Lactamase-Inhibitor Protein Binding Affinities: Detection of Hydrolysis of ß-Lactam Substrate by TEM-1 Enzyme and Inhibitor Binding Using The FlexStation® 3 Microplate Reader
Bacterial resistance to ß-lactam drugs through the production of class A ß-lactamase enzymes is an increasing concern in the medical community.1 ß-lactamase inhibitory protein (BLIP)…
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Quantitation of dsDNA samples using the SpectraMax Quant dsDNA Assay Kits
Quantitation of dsDNA samples using the SpectraMax Quant dsDNA Assay Kits
As different types of test samples may contain different concentrations of DNA, selection of an appropriate fluorescent reagent with the required sensitivity and detection range is…
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Noninvasive measurement of fluorescent proteins in live cells
Noninvasive measurement of fluorescent proteins in live cells
Fluorescent proteins have become enormously popular as tools for monitoring biological events in vivo. Green fluorescent protein (GFP) from the jellyfish Aequorea victoria was the first…
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IMAP phosphodiesterase assays on SpectraMax Multi-Mode Microplate Readers
IMAP phosphodiesterase assays on SpectraMax Multi-Mode Microplate Readers
Cyclic nucleotide phosphodiesterases (PDEs) are a group of enzymes that degrade the phosphodiester bond of cAMP and cGMP, second messengers that are involved in a variety of biological…
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Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader
Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader
The CellTiter-Glo assay from Promega uses luciferase enzyme, which requires ATP in order to generate light. The luminescent signal produced in the assay depends on the amount of ATP in the…
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Detection and quantitation of protein with ScanLater Western Blot Detection System
Detection and quantitation of protein with ScanLater Western Blot Detection System
Protein detection is important for pharmaceutical and clinical research today, and western blots are among the most common methods employed for this purpose. Various techniques are used to…