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Norgen’s FBS Exosome Depletion Kits (Slurry Format) constitute an all-in-one system for the depletion of cow’s exosomes from FBS prior to using it as a growth supplement in your culture medium.

• Efficient depletion of cow’s exosomes from Fetal Bovine Serum
• Deplete exosome-sized vesicles from versatile FBS volumes of up to 280 mL
• No protease treatment required
• No time-consuming ultracentrifugation
• No precipitation reagents required
• No overnight incubation required
• Depleted FBS has no detectable cow’s miRNA
• The depleted FBS provides the same cellular growth rates as the standard FBS

FBS Exosome Depletion Kit (Slurry Format) 61100

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FBS Exosome Depletion Kit (Column Format) Figure 2

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Preparing protein samples for mass spectrometry often involves tedious, multi-step protocols that increase the risk of sample loss and contamination. The ProteoSpin™ On-Column Proteolytic Digestion Kit provides a revolutionary solution by integrating protein purification, digestion, and peptide recovery into a single, streamlined spin-column procedure. This approach eliminates the need for time-consuming overnight dialyses or complex precipitation steps, ensuring that researchers can transition from raw protein samples to MS-ready peptides in a fraction of the traditional time.

The system utilises advanced ion-exchange chromatography within a convenient spin-column format. Proteins are first bound to the column matrix, where contaminants such as salts, detergents, and buffers are washed away. Proteolytic enzymes (such as Trypsin) are then introduced directly onto the column for efficient digestion. Because the digestion occurs in a concentrated environment on the matrix, the reaction kinetics are highly favourable. Following digestion, the resulting peptides are eluted in a small volume of MS-compatible buffer, ready for immediate downstream analysis without further clean-up.

Key Benefits:

• Unified Workflow: Purification and digestion in one. Combines sample clean-up and enzymatic digestion into a single protocol, significantly reducing manual handling time.
• High Peptide Recovery: Maximise your data. The on-column format minimises sample transfer steps, ensuring maximum recovery of even low-abundance peptides for sensitive LC/MS applications.
• Rapid Processing: Results in hours, not days. Accelerated digestion kinetics and efficient elution allow for the completion of the entire process in significantly less time than traditional in-solution methods.
• Superior Purity: Optimised for MS. Effectively removes common MS-interfering substances, including SDS, Triton™ X-100, and high salt concentrations, leading to cleaner spectra and better base calling.

Technical Insights

The ProteoSpin™ technology is engineered to handle a wide range of protein concentrations. By immobilising the protein on the column, the effective concentration of the substrate is increased, allowing for efficient digestion even when using lower ratios of expensive enzymes like Trypsin. The kit is compatible with various proteolytic enzymes, providing flexibility for different proteomic mapping requirements.

Specifications

• Sample Type: Purified proteins or complex lysates
• Format: Spin Column (Ion-exchange chromatography)
• Processing Time: Approximately 3–4 hours (including digestion)
• Downstream Compatibility: LC-MS/MS, MALDI-TOF, HPLC
• Storage Conditions: Room temperature (Enzymes may require -20°C if supplied)

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The Detergent-Free Total Protein Isolation Kit maintains high protein recovery and yields Proteins that are 100% detergent-free. Purification is based on using proprietary resin together with Lysis Solution, followed by protein filtration using a filter column (provided). The purified Proteins can be used in a number of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, NMR spectroscopy and more.

Each Lysis Tube is able to process up to 50 mg of tissue, 1010 bacterial cells, 109 yeast cells or 107 mammalian cells Preparation time for 12 samples is less than 10 minutes.

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Homologous recombination is a critical cellular process, and measuring its efficiency is essential for understanding DNA repair mechanisms, gene targeting, and genetic stability. Traditional methods of assessing recombination efficiency often rely on labour-intensive protein expression and staining techniques, which limit scalability and extend experimental timelines. The Homologous Recombination Assay Kit provides a rapid, robust, and highly sensitive solution for precisely quantifying recombination events in both bacterial and mammalian cells, eliminating the bottleneck of phenotypic screening.

The assay leverages a co-transformation strategy using two distinct plasmids (dl-1 and dl-2), each containing different defective mutations in the lacZα coding region. When these plasmids are co-transfected into a host cell, intermolecular homologous recombination can generate a functional lacZα cassette. Recombination efficiency is then accurately quantified using qPCR, facilitated by the provided assay and universal primer mixtures designed to amplify only the recombined product. This PCR-based approach significantly enhances sensitivity and streamlines the workflow, making it ideal for high-throughput screening applications.

Key Benefits:

• Rapid and Sensitive Quantification: PCR-based precision. Utilises qPCR to deliver highly sensitive and quantitative measurement of homologous recombination events, eliminating the need for slow, traditional staining methods.
• Broad Compatibility: For multiple cell types. Optimised for evaluating recombination efficiency in both prokaryotic (bacterial) and eukaryotic (mammalian) cell models, offering versatile experimental applications.
• Comprehensive Kit: All essential primers included. Comes complete with specific assay primers and universal primer mixtures, ensuring targeted amplification of only the successful homologous recombination product.
• High-Throughput Ready: Streamline your screening. The efficient qPCR-based protocol is perfectly suited for high-throughput genetic screening, saving valuable laboratory time and resources.

Technical Insights

Figure 1: Schematic Representation of Homologous Recombination Assay

The lacZα coding region of the positive control plasmid is mutated to generate two specific plasmids (dl-1 and dl-2) with distinct mutations. These two defective plasmids can form a functional lacZα cassette through intermolecular homologous recombination. When transformed into bacterial cells, defective plasmids form white colonies, while plasmids that have successfully undergone homologous recombination to generate functional lacZα form blue colonies. Efficiency can be determined quantitatively via qPCR using the supplied primer mixtures, or qualitatively by calculating the percentage of blue colonies in a traditional blue/white screening assay.

Figure 2: Real-time PCR Amplification of Plasmid DNA Isolated from HEK 293 Cells.

After transfection with the supplied plasmids genomic DNA was isolated from each well of a 6-well plate using Norgen’s RNA/DNA/Protein Purification Kit. Fifty nanograms of the positive control plasmid was transfected into each well of the HEK 293 cells, and the total DNA was isolated 12 h later. For the recombinant samples, 25 ng of dl-1 plus 25 ng of dl-2 were transfected and then isolated. For the negative controls, 50 ng of dl-1 or 50 ng of dl-2 were transfected and then isolated. The experiment was carried out in duplicate to avoid the reading error of the PCR machine.

Specifications

• Kit Capacity: 200 transformations or 24 transfections
• dl-1 Plasmid: 12 μg (50 ng/μL)
• dl-2 Plasmid: 12 μg (50 ng/μL)
• Positive Control Plasmid: 5 μg (50 ng/μL)
• Assays Primers Mixture: 400 μL
• Universal Primers Mixture: 400 μL
• Storage Conditions: All components should be tightly sealed and stored at -20°C or lower.
• Shelf Life: Stable for 5 years after the date of shipment.
• Intended Use: For research use only; not intended for in vitro diagnostics.

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