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Application Notes

  • Label-free detection of transporter activity via GpcR signalling in living cells: A case for SLC29A1, the equilibrative nucleoside transporter 1

    Transporters are important therapeutic but yet understudied targets due to lack of available assays. Here we describe a novel label-free, whole-cell method for the functional assessment of Solute Carrier (SLC) inhibitors. As many SLC substrates are also ligands for G protein-coupled receptors (GPCRs), transporter inhibition may affect GPCR signalling due to a change in extracellular concentration of the substrate/ligand, which can be monitored by an impedance-based label-free assay.
  • CAR-T Cells for the Treatment of Blood and Solid Tumors

    The development of CAR-T cells began in the late 1980s-1990s and the technologies that ultimately led to their creation were first developed by immunologists Michel Sadelain and Zelig Eshhar. At the time the first T cell engineering was taking place, most scientists did not believe that T cells would have much efficacy in the fight against cancer. Little could they have predicted that two decades later, CAR-T cells would be an FDA-approved treatment for adult and childhood cancer.
  • Generating Clonal GFP Cell Lines

    Generating a clonal GFP cell line typically requires first sorting GFP positive cells with a cell sorter. Then GFP positive cells are cultured for several days to recover from the high pressure damage caused by the cell sorter. Clonal GFP cells are finally obtained by manual limited dilution at 1 cell per well. In this application note, Namocell Single Cell Dispenser was used to directly isolate single GFP positive cells into 96-well cell culture plate. Due to the low pressure of the Namocell Single Cell Dispenser, clonal GFP cell lines can be generated in a single step.
  • Single B Cell Genomics

    By use of a Namo Single Cell Dispenser, activated single B cells were isolated. Single B cell cDNA was suitable for both downstream single cell transcriptome library generation and PCR of the full length of immunoglobulin heavy and light chain variable domains.
  • Minimal Cell Input

    Obtaining a single cell is a critical step for single-cell transcriptomics, proteomics, and genomics. Traditional cell sorters have provided a powerful tool for sorting cells. However, it typically requires a large amount of cells. In this application note, Namocell Single Cell Dispenser was used to evaluate the single cell isolation efficiency of 100 minimal cell input with an adherent and non-adherent cell line. Complete your details and receive a copy of the application note in your inbox.
  • Single Nuclei Dispensing

    Using a Namocell Single Cell Dispenser, single nuclei were isolated and verified under a fluorescent microscope. Isolated single nuclei are suitable for single-nucleus RNA sequencing. Complete your details and receive a copy of the application note in your inbox.
  • NEBNext Direct® Custom Ready Panel technology analyzed using NextGENe®

    With targeted panels becoming an excellent alternative for whole exome and genome Next Generation Sequencing (NGS) there is an increasing need for highly flexible and optimized custom panels. The NEBNext Direct technology in conjunction with NextGENe software represents a high-quality solution for generation and analysis of sequencing data for identification sequence variants present in a user-friendly manner.

  • Medimachine II for Lymph node dissociation and subsequent cytometric analysis

    Standardization in tissue disaggregation minimizes differences in cells by fresh and frozen tissue and focuses relevant variations on both surface marker detection and functional parameters

  • Viral Vaccine Development Gets Major Boost From xCELLigence® RTCA

    Using xCELLigence Real-Time Cell Analysis (RTCA), Sanofi Pasteur scientists have developed an assay for accurate viral titer determination that is 5-times less labor intensive and 3.5-times cheaper than conventional methods.

  • Scientists at Roche Developed an Impedance-Based Contraction Assay Using Human Primary Smooth Muscle Cells and Fibroblasts

    The measurement of contractility of Bronchial smooth muscle cells (BSMCs) and airway fibroblasts are important to study the pathogenesis of allergy/asthma and the mechanism of action of anti-allergic/asthmatic drugs. The conventional gel contraction assay is frequently used to study both fibroblasts and smooth muscle cells in vitro, but it is extremely laborious and low throughput.

  • Upgrade Your T-Cell Marker Research With Bethyl Antibodies

    T-Cell Marker Antibodies: Advancing Cancer Immunotherapy Breakthroughs

  • Mechanical DNA Fragmentation with the Q800R2 Sonicator

    The Qsonica Q800R2 Sonicator offers an alternative method for mechanical DNA fragmentation for Illumina TruSeq® Library Preparation Kits.

  • Restriction Enzymes for Droplet Digital PCR (ddPCR)

    NEB has evaluated a number of restriction enzymes for simple genomic DNA fragmentation in droplet digital PCR assays. This list is not meant to be exhaustive, but provides a starting point for validated enzymes with some simple usage guidelines. Each restriction enzyme listed below was successfully used for a ddPCR digest directly in the Bio-Rad® Droplet Digital SuperMix.

  • Analysis of chromosomal aberrations and recombination by allelic bias in RNA-Seq

    Read this publication in Nature Communications about the successful RNA purification for downstream RNA sequencing with the NucleoSpin RNA Plus Kit.

  • ACEA NovoCyte Flow Cytometer with 561nm Laser

    The expansion of flow cytometry applications from cellular analysis to molecular and genomic analysis, and the increase in availability of monoclonal antibodies and fluorochromes have magnified the demand for flow cytometers with expanded fluorescence detection channels. ACEA NovoCyte flow cytometers are configured with 561nm laser to meet the needs of multicolor analysis, broadening application capabilities in both basic research and clinical studies.

  • Characterize Immune Checkpoint Proteins and T Cell Exhaustion Using Multiplex IHC

    Advances in immuno-oncology have successfully led to novel cancer therapeutics with favorable patient responses that are more durable than conventional cytotoxic chemotherapy (1). However, not all patients respond to immunotherapy; therefore investigators are trying to identify clinically relevant biomarkers with the goal of developing therapeutics based on personalized medicine (2,3).

  • NucleoBond Endotoxin Free Plasmid DNA in CRISPR/Cas9 and lentiviral packaging/cell culture research

    Forward genetic screens are powerful tools for the unbiased discovery and functional characterization of specific genetic elements associated with a phenotype of interest. Recently, the RNA-guided endonuclease Cas9 from the microbial CRISPR (clustered regularly interspaced short palindromic repeats) immune system has been adapted for genome-scale screening by combining Cas9 with pooled guide RNA libraries. Here we describe a protocol for genome-scale knockout and transcriptional activation screening using the CRISPR-Cas9 system. 
    Read about the advantage of using NucleoBond Endotoxin Free Plasmid DNA from MACHEREY-NAGEL to avoid endotoxicity in virus production and mammalian cell culture. 

  • Hard-to-Lyse Samples

    Struggling with DNA isolation from hard-to-lyse samples? Up to 60% more DNA compared to standard extraction methods.

  • Chromatin / DNA Shearing with Qsonica Sonicators

    Qsonica offers several devices routinely used for shearing Chromatin and DNA. The Q800R system is our most advanced option and offers the highest throughput. In addition, the Q700 and Q125 are very popular and effective alternatives.

  • Nanoparticle Dispersion with Qsonica Sonicators

    Probe sonication is highly effective for processing nanomaterials (carbon nanotubes, graphene, inks, metal oxides, etc.). Sonicators have become the industry standard for different applications.

  • Cell Disruption and Homogenization with Qsonica Sonicators

    Cell lysis, tissue disruption and homogenization are common Sonicator applications. The ultrasonic energy output of each Sonicator model is adjustable and sonication parameters can be optimized according to your process requirements.

  • NEBuilder HiFi DNA Assembly versus Gibson Assembly

    This application note describes how to assembly multiple fragments and have 100% correct clones.

  • NEB Restriction Enzymes - Inside the Numbers

    After nearly 40 years of offering restriction enzymes to the research community, New England Biolabs (NEB) has earned the reputation of being a leader in enzyme technologies. NEB offers scientists 276 restriction enzymes, the largest selection in the industry.

  • Xtra Quick & Simple DNA Isolation

    A PhD student working at the Department of Immunology at the University Medical Center Utrecht shares her positive experience with the NucleoBond Xtra Midi Kit from MACHEREY-NAGEL

  • Cellular Imaging

    Visualization of Wnt receptor internalization by SNAP-tag technology from New England Biolabs (NEB)

  • Epigenetic landscape of proximal tubular epithelial cells using ChIPseq

    The epigenetic status of genes can be assayed by chromatin immunoprecipitation (ChIP). ChIP is a powerful technique to study DNA-protein interactions. To explore the genome-wide binding of transcription factors and the epigenetic landscape ChIP can be combined with sequencing (ChIPseq). This technique enables exploration of the epigenetic state in a global fashion

  • Gibson Assembly

    In this application note we describe the use of the Gibson Assembly™ Master Mix from New England BioLabs (NEB) together with gBlocks™ Gene Fragments from Integrated DNA Technologies (1) for the creation of a basic cloning vector for IP3R research.

  • How specific is your IHC antibody?

    Validating an antibody for immunohistochemistry (IHC) use involves more than just observing a signal in cells.

  • Q5 High Fidelity DNA Polymerase

    Marjorie Mercier, a scientist in Brussels, shares her positive experience with Q5 High-Fidelity DNA Polymerase for the amplification of the IgM variable region.

  • Q5 High Fidelity DNA Polymerase

    Colette Duez of the Center of Protein Engineering/ULg shares her positive experience with Q5 High-Fidelity DNA Polymerase for the amplification of a GC-rich (73%) insert of 1.4 kb from bacterial origin.

  • Q5 High Fidelity DNA Polymerase

    Mrs. Evy Vanderheyden, working at the VIB/KULeuven Molecular Microbiology and Biotechnology department shares her positive experience with Q5 High-Fidelity DNA Polymerase for the robust amplification of genomic yeast DNA.

  • Site-Directed Mutagenesis (SDM)

    Improved methods for site-directed mutagenesis using Gibson Assembly Master Mix from New England Bioloabs

  • Chromatin Immunoprecipitation

    SimpleChIP® in Literature: A synopsis of a recent publication by Tomas J. Bos, Ph.D., Department of Hematology and Immunology, Vrije Universiteit Brussel, Brussels, Belgium

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