Agilent Manuals

This Application Note describes how the Agilent 2100 bioanalyzer and the cell fluorescence LabChip® kit can be used to determine the efficiency of transfection of mammalian cells using green fluorescent protein (GFP) as a reporter molecule by on-chip staining. Transfection of CHO-K1 cells with an expression vector encoding GFP was performed and cells were stained on-chip with the live-dye CBNF (carboxynaphthofluorescein). The histogram quality and the percent transfected cells determined are in good agreement with data obtained using a conventional flow cytometer. Detailed protocols and reagent recommendations for analyzing transfection reactions are provided. Speed of the on-chip staining procedure, high reproducibility of the chip results, low cell consumption and ease-of-use are advantages the Agilent 2100 bioanalyzer offers for monitoring transfection efficiency using GFP.

This Application Note describes a new protocol for detecting cell surface protein targets with the Agilent 2100 bioanalyzer and the cell fluorescence LabChip® kit. The specific advantage of an on-chip staining reaction is the amount of time, cells and reagents saved. CD3 on-chip staining on Jurkat cells as well as CD80 (B7-1) and CD86 (B7-2) on- chip staining on 293 cells stably transfected with a CD80 or CD86 expression plasmid were performed. The histogram quality and the percentage of stained cells counted with the microfluidic system are in good agreement with data obtained using a conventional flow cytome- ter. Detailed protocols and reagent recommendations for on-chip stain- ing of cell surface proteins are provided. The new procedure requires 17 fold less cells per sample analysis compared to the conventional staining procedure, while reducing antibody reagent costs by a factor of 80. The high reproducibility of the chip results, very low cell and reagent consumption as well as speed and ease-of-use are advantages the Agilent 2100 bioanalyzer offers for monitoring cell surface protein targets by on-chip antibody staining.

This Application Note describes how the Agilent 2100 bioanalyzer and the Cell Fluorescence LabChip® kit can be used to detect cell surface and intracellular protein targets by antibody staining. CD3 staining on Jurkat cells as well as CD4 staining on CCRF-CEM cells and intracellular glucocorticoid receptor (GR) staining of H4 rat hepatocytes were performed. Histogram quality and the percentage of stained cells counted with the microfluidic system are in good agreement with data obtained with a conventional flow cytometer. Detailed protocols and reagent recommendations for staining cell surface and intracellular protein targets are given. The high reproducibility of the chip results, low cell consumption and ease-of-use are advantages of the compact bioanalyzer system for monitoring cell surface and intracellular protein target expression by antibody staining.

This Application Note describes a simplified and fast protocol for the detection of apoptotic cells with the Agilent 2100 bioanalyzer and the cell fluorescence LabChip®. The specific advantage of the simplified protocol is the amount of time saved and its ease-of-use. The note describes how the Agilent 2100 bioanalyzer can be used to analyze apoptotic cell samples. Annexin V staining of live cells is demonstrated and the data are compared to that obtained with a conventional flow cytometer. Histogram quality of the microfluidic system is comparable to that of a conventional flow cytometer. Detailed protocols and reagent recommendations for staining of cells are provided. The high reproducibility of the chip results, low cell and reagent consumption, the speed and the ease of use are advantages the Agilent 2100 bioanalyzer offers for monitoring apoptotic cells by annexinV staining.

This application note describes how the Agilent 2100 bioanalyzer can be used in combination with Standardized RT (StaRT)-PCR reagents from Gene Express National Enterprises (G.E.N.E.), Inc. (Huntsville, AL, USA) to obtain highly reproducible, standardized gene expression measurement with RT-PCR. The internal standards used in StaRT-PCR control for variable loading of PCR products into electrophoresis chan- nel, and allow quantitative gene expression measurement at the end- point of PCR, without real-time analysis. Gene expression measure- ment of 15 genes was more reproducible with the Agilent 2100 bioana- lyzer than with the ABI Prism 310 Genetic Analyzer or agarose gel elec- trophoresis methods. The mean coefficient of variance (CV) for these three methods was 0.26, 0.39 and 0.50, respectively.

This article describes the analysis of Cy5-labeled cRNAs and cDNAs using the Agilent 2100 Bioanalyzer and the RNA 6000 LabChip® kit. It discusses various methods for detecting and quantifying gene expression levels, as well as the use of fluorescently labeled probes in microarray applications. The importance of using high-quality RNA for synthesizing labeled cDNAs or cRNAs is emphasized.

Agilent 2100 Bioanalyzer is an integrated, inexpensive, compliant solution for the analysis of nucleic acids, proteins, or cells. The system can easily be used for the quality control and analysis of antibodies, protein pharmaceuticals, and other biomolecules such as RNA and DNA.

This document introduces the characteristics of Agilent 1290 Infinity LC and its complementary sensitivity with MS detection.

Apoptosis detection by annexin V and active caspase-3 with the Agilent 2100 bioanalyzer

This application note describes the separation of 4 mycotoxins using HPLC with the Agilent Pursuit XRs Ultra 2.8 Diphenyl column. The elution was isocratic, the flow rate was 0.4 mL/min, the temperature was 40 °C, and the detection wavelength was 236 nm.

This article introduces a target enrichment technology based on Agilent DNA microarrays, which can be used to resequence multiple discrete genomic regions of interest.

This document describes how the Agilent High Sensitivity DNA Kit and the Agilent SureSelect Target Enrichment Platform can improve sample quality for target enrichment and next-generation sequencing. By utilizing improved detection sensitivity, the number of library PCR cycles can be reduced, eliminating amplification bias and significantly improving the quality and accuracy of NGS data.

Direct measurement of metallic impurities in 20% ammonium hydroxide by Agilent 7700s ICP-MS

This application note investigates the ability of the Agilent Fish ID solution to detect mammalian or avian DNA using meat samples.

The application note shows the analysis of Tetrahydrothiophene (THT) in a Natural Gas matrix using the Agilent 490 Micro GC.