Agilent Manuals

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 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 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 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 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 investigates the various techniques for RNA quantitation, including a comparison of the Agilent 2100 bioanalyzer with UV spectrometry and the Ribogreen® quantitation system to access the linear dynamic range, reproducibility, and the effects of contaminants for each quantitation method.

This article describes the analysis of nucleotides using an Agilent Polaris C18-A column. The HPLC separation of ten nucleotides was achieved in 24 minutes. The column has a particle size of 5 μm and is suitable for pharmaceutical applications.

Introduces the separation of four metanephrines in urine with an Agilent Polaris C18-A column

Agilent RNA 6000 Pico LabChip® kit can be used to control the quality of RNA in laser microdissected tissue samples

A robust method for detection of THC and its metabolites in blood has been developed using SPE extraction and GC/MS/MS with backflushing. The dynamic range of quantification was 0.1 to 50 ng/mL for THC and 11-OH-THC, and 1 to 100 ng/mL for THCA, with a run time of 6 minutes and a cycle time of 8 minutes.

This Application Note describes a method for the fast analysis of chemotherapy drugs using Agilent Polaris and Agilent Pursuit columns. The method allows for the differentiation of cancer drugs in less than three minutes and can be used to assess the quality of drug preparations. The development of this method is a major step in improving the quality of drug delivery for cancer patients.

This document describes the method of using Agilent ResiPore columns and gel permeation chromatography to analyze phenolic resins

This document introduces the application of Agilent OligoPore and gel permeation chromatography to analyze silane oligomers, which are generated by the controlled hydrolysis of silanes under acidic conditions, and the silane oligomers are then analyzed by Agilent OligoPore columns.

This document describes the use of Agilent OligoPore and gel permeation chromatography for fingerprinting epoxy resins. The distribution of oligomers in the resin determines its physical properties, such as curing time and mechanical strength. The analysis of oligomer distribution using gel permeation chromatography allows for the fingerprinting of different grades of resin, which is crucial for quality and process control. The authors demonstrate the analysis using two commercial epoxy resins, Epikote 828 and Epikote 1001. Epikote 828 is dominated by a single oligomer peak, while Epikote 1001 contains multiple oligomers with increasing molecular weight. This method is of significant importance for resin quality and process control.