| Structural Biology Applications #15 |
Structural biology is an important part of Life Science Research focusing on the molecular structure and formation of biological macromolecules ( e.g. proteins, consisting of amino acids, DNA and RNA consisting of nucleotides and cell membranes consisting of lipids). Structural biology is of key interest to biologists as macromolecules perform a majority of cell functions, which are determined primarily by the unique three-dimensional formation of each molecule. This architecture, determined by the “tertiary structure”, depends on the linear sequence considered the “primary structure” of each molecule. The understanding of the tertiary (3D) structure of a molecule is a basic prerequisite for understanding function… |
| Best Practice for Protein Quantifications and Antibody Quantifications #14 |
Protein concentrations are frequently used in life science research. Although there are several assays available, care should be taken to select the optimum assay or method for the particular sample type. The decision on which assay or method to use is typically based on convenience, quantity and purity of protein available. In this technical note, we provide best practice advice for using the NanoPhotometer® to perform small volume protein quantification at 280 nm (Protein UV280) and also point to protein assays including Bradford, BCA, etc. for the indirect determination of the protein concentrations via a chemical binding reaction… |
| The Limitations of DNA Quantification Using DNA Binding Fluorescent Dyes #13 |
For the quantification and the quality control of a nucleic acid sample, a UV/Vis spectroscopic reading at 260 nm is recommended to obtain concentration and purity of the specimen simultaneously (260/280 and 260/230 ratios) and without the need for tedious and error prone preparation steps or expensive consumables. All types of nucleic acids typically show a concentration absorbance peak around 260 nm, which can make it challenging to reliably distinguish between DNA and other types of nucleic acids using direct UV/Vis spectroscopy.
Most extraction protocols used for daily routine applications are optimized to remove all unwanted fragments and nucleic acid types with appropriate purification steps. However, in case of potential insufficient purification or in rare cases determined by the nature of the extraction, a fluorescence based analysis is recommended to determine relatively accurate dsDNA concentration readings (e.g. dsDNA from FFPE samples)… |
| Best Practice for Nucleic Acid Quantifications #12 |
Nucleic Acid quantification is an important step for many different life science protocols including those implemented for NGS, qPCR and several others. The NanoPhotometer® offers preprogrammed Nucleic Acid methods with options to measure all types of nucleic acids including dsDNA, ssDNA, RNA, miRNA, and Oligos, as well as the option to enter sequences for miRNAs and Oligos as a custom defined extinction coefficient for other types of nucleic acids. It is also possible to measure fluorescent labeled nucleic acids, e.g. as a quality control step during micro array workflows, via the absorbance maxima of the dye. Dye concentration and the frequency of incorporation are automatically calculated and displayed… |
| Cleaning of the NanoPhotometer® Microvolume Measurement Head for Nucleic Acid, Protein and Antibody Quantification #11 |
The microvolume measurement head of the NanoPhotometer® consists of scratch resistant quartz surfaces making it very easy to clean. In comparison to other technologies that are utilizing stainless steel, the inert NanoPhotometer® microvolume measurement head never needs to be reconditioned over the entire lifetime – even after the application of highly concentrated protein samples… |
| Sample Control™ Technology for Nucleic Acid and Protein Quantification #10 |
Measurements in a drop have become state-of-the-art and very simple to do over the past two decades, however, there are some areas where sample handling errors and/or contaminations may impact the quality of measurement results. With NanoPhotometer® Sample Control™, we have created a valuable tool to cover main areas of concern (air bubbles, insufficient cleaning and/or unwanted residuals, etc.) ensuring that researchers always can trust their readings… |
| Bradford Assay in Microvolume #9 |
Rapid and accurate estimation of protein concentration is essential in laboratory practice. Depending on the protein sample and buffers utilized, the optimal spectrophotometrical method should be performed. Direct protein UV measurements at 280 nm require pure proteins solved in buffers without absorbance interference at 280 nm. If these conditions are unable to be met, it is recommended to use a protein assay: Bradford, Biuret, BCA or Lowry.
The optimal colorimetric assay should be chosen depending on the linear range of the assay and the interference of buffer components with the assay reagents. For more details please refer to the manufacturer’s directions… |
| OD600 | Cell Density, Bacterial Growth, Yeast Growth #8 |
Optical Density (OD) measurements of microbial and cell growth are one of the most common methods used in a microbiology lab. Some of the main applications are the determination of the optimal time at which to harvest, the determination of the optimal time to induce a culture when running a protein expression protocol or the monitoring of cloning procedures.
The growth of cells, bacteria or yeast (cell density, bacterial growth, yeast growth) in liquid culture media is commonly controlled by measuring the optical density at 600 nm (OD600). OD600 measurements are typically used to determine the stage of growth of a bacterial culture, these measurements help ensure that cells are harvested at an optimum point that corresponds to an appropriate density of live cells… |
| UV Vis Spectrophotometer | Flexible Unit Control #7 |
The NanoPhotometer® Family offers a unique range of nanovolume and/or cuvette UV/VIS spectrophotometers with flexible control options. All NanoPhotometer® models can be accessed and controlled by a variety of control devices. In addition, a built-in touch screen option is available for independent stand-alone operation… |
| UV Vis Spectrophotometer Data Management #6 |
The NanoPhotometer® family of UV/Vis spectrophotometers provides a well-structured data management portfolio to store different kinds of file formats in dedicated folders on different devices. |
| Protein UV Performance Data | How to Measure Protein UV Concentrations | Protein UV Quantification #5 |
Proteins, the dynamic parts of living cells, are involved in nearly all metabolic pathways of cells (e.g. enzymatic reactions, immune protection, and cellular response). Protein UV Spectrophotometers are frequently used for the measurement of Protein UV concentrations in life science research. Although there are many assays available, care should be taken to select the optimal assay for the particular sample type. The decision on which assay to use is typically based on convenience, quantity and purity of protein sample available. It is also possible to measure labeled proteins via the absorbance maxima of the dye. Dye concentration and the frequency of incorporation are calculated and displayed automatically… |
| Nucleic Acids Performance Data | How to Measure Nucleic Acid Concentrations | Nucleic Acid Quantification #4 |
Nucleic Acid quantification is an important step in many different life science protocols including those implemented for NGS, qPCR, and many other areas. The Nucleic Acid Spectrophotometer, NanoPhotometer® by Implen, offers a preprogrammed Nucleic Acid method with options to measure all types of nucleic acids including dsDNA, ssDNA, RNA, miRNA, Oligo, options to enter sequences for miRNAs and Oligos as well as a custom-defined extinction coefficient for other types of nucleic acids. It is also possible to measure labeled nucleic acids via the absorbance maxima of the dye. Dye concentration and the frequency of incorporation are calculated and displayed automatically. |
| UV-Vis Spectrophotometer Blank Control #3 |
The blank measurement as the first step for all photometric measurements is the most critical part for accurate and reproducible sample readings. It is a prerequisite for all photometric measurements.
This Technical Note gives useful information about the blank solution, blank quality and information about the unique Blank Control™ feature in the NPOS software. |
| UV/Vis Spectrophotometer Sample Compression Technology™ #2 |
The patented Sample Compression Technology™ provides unmatched precision and accuracy with reliable measurement geometry even for challenging samples including proteins and products in volatile solvents. This technology is especially designed for ultra-low sample volumes of 0.3 µl to 2 µl.
Principle of Small Volume Measurements with a UV/Vis Spectrophotometer… |
| UV/Vis Spectrophotometer True Path Technology™ #1 |
The NanoPhotometer® NP80/N60/N50 is designed to provide accurate and reliable measurements for ultra-low volume samples of 0.3 µl to 2 µl. No recalibration is needed for the entire lifetime of the instrument. The unique True Path Technology™ is the backbone of this important feature, which creates a calibrated optical environment that does not change over time.
Principle of Small Volume Measurements with a UV Vis Spectrophotometer… |
