Agilent Manuals

This document describes Agilent's dual-mode Gene Expression Microarray platform and the method for gene expression analysis using RNA isolated from human whole blood. The platform combines the ease and simplicity of one-color experimental design with the capability of two-color sensitivity for detection of small gene expression changes. Agilent's platform now includes a method for the analysis of differential gene expression from human whole blood, which is compatible with the PAXgene™ Blood RNA System. The PAXgene Blood RNA System consists of a blood collection tube and nucleic acid purification kit for the collection, storage, and transport of blood and stabilization of intracellular RNA for subsequent isolation and purification. Combined with the PAXgene System, Agilent's amplification and labeling technology, in conjunction with the high sensitivity and specificity of 60-mer oligonucleotide microarrays, offer a total solution for gene expression in whole blood without the need for additional steps or kits to remove globin transcripts. Titration experiments using Agilent RNA spike-in control targets demonstrate the sensitivity of detection.

This article introduces an integrated approach to improve sequence coverage and protein identification by combining liquid chromatography-matrix-assisted laser desorption ionization mass spectrometry (LC-MALDI MS/MS) and nanoflow liquid chromatography-tandem mass spectrometry (nano-LC/MS/MS). The coupling of RP-HPLC with MALDI via a MALDI spotting device provides an automated and robust solution for analyzing medium complexity samples using MALDI. The experimental results show that this integrated approach can improve protein coverage for complex protein samples and facilitate protein identification.

Confirmation of THC in Oral Fluids Using High-Resolution 2-D GC/MS

This document introduces a method for isolating total RNA samples using the Agilent Total RNA Isolation Kit and the Agilent DNase Accessory Kit, which can effectively reduce the genomic DNA contamination in total RNA samples.

Efficient Removal of Transfected Plasmid DNA from Total Cellular RNA Prepared with Agilent Total RNA Isolation Mini Kit Application

A protocol for the isolation of high-purity RNA from gram-positive and gram-negative bacteria was developed for use with the Agilent Total RNA Isolation Mini Kit.

This study describes a method based on Agilent Multiple Affinity Removal System for removing high-abundance proteins from monkey plasma, which can increase the sensitivity of detecting low-abundance proteins.

This paper introduces a new multidimensional separation technique to reduce the complexity of complex serum samples. The technique uses a high-capacity multiple affinity removal column to specifically remove six high-abundance proteins (albumin, IgG, transferrin, haptoglobin, IgA, and anti-trypsin), and then uses a macro-porous reverse-phase C18 (mRP-C18) column to fractionate the depleted serum. Under the optimized mRP-HPLC condition, protein recoveries for depleted serum samples were greater than 99%. Reproducibility among patient samples was excellent based on the UV trace overlays. The mRP-C18 fractionated serum samples were further analyzed by 1D-PAGE and 2D-LC-MS/MS, the results showed that the technique can successfully reduce the complexity of serum samples and improve the detection sensitivity of low-abundance proteins.

The document introduces the Agilent Technologies' Multiple Affinity Removal System, which combines the specificity of antibody-antigen recognition with the efficiency of standard liquid chromatography instrumentation. This system effectively removes multiple proteins from human serum, including albumin, transferrin, IgG, IgA, haptoglobin, and antitrypsin, in a single step with high specificity and reproducibility. The document also demonstrates several methods to optimize column binding capacities, such as coupling two identical columns to increase capacity, using a custom column with dimensions of 10x100mm to enhance capacity, and achieving optimal binding capacity by injecting samples at a low flow rate.

The search for biomarkers in proteomics research continues to be a critical bottleneck in the development of new drugs as well as in the study of disease. This paper discusses the limitations of traditional protein analysis methods and proposes the use of specific interaction media for the selective removal of high-abundant proteins.

This application note describes the application of the Low RNA Input Fluorescent Linear Amplification kit for the use of poly (A)+ RNA or total RNA samples. It shows that micrograms of cRNA targets can be generated within 6 hours, using as little as 10 ng of poly (A)+ RNA, or 50 ng of total RNA sample input. The resulting cRNA targets were of high quality and per- formed reproducibly and robustly on Agilent's oligonu- cleotide microarrays. Furthermore, the log ratios of array results from poly (A)+ RNA sample input correlated with that from total RNA input. The easy-to-use Low RNA Input Fluorescent Linear Amplification kit can be applied to a wide range of samples for gene expression analysis.

Fast and Ultra-fast Analysis with the Agilent 1200 Series Rapid Resolution LC System Compared to a Conventional Agilent 1100 Series LC System Using Sub 2-µm Particle Columns

This Application Note describes a method for studying noncovalent protein interactions using the HPLC-Chip/MS system coupled with an ESI-TOF mass spectrometer. The application note studies apomyoglobin and myoglobin protein complexes. The results show that ESI/MS is an efficient and accurate method for studying noncovalent protein interactions.

Determination of Toxic Elements in Traditional Chinese Medicine Using Inductively Coupled Plasma Mass Spectrometry

This article describes the analysis of a complex natural product extract from ginseng. By using LC/MS technology, it is possible to differentiate between different species of ginseng plants and authenticate ginseng products.