cell separation using pro5 pentamers and magnetic beads proimmune

Analysis of antigen‐specific CD8 + T cells using class I pentamers HLA‐A*0201 pentamers (ProImmune, Oxford, UK) loaded with the autoantigenic epitopes of choice, positive control viral epitope(s) and negative control epitope. Materials required. Cell sample, e.g. blood sample (RBC‐depleted), PBMCs or T cell line. • Pro5 recombinant MHC pentamer conjugated to the Magnetic Bead Cell Separation - ProImmune - Mastering, Cell Separation using Pro5 Pentamers and Magnetic Beads In addition to detecting antigen-specific T cells, Pro5 MHC Class I Pentamers can be used in conjunction with magnetic beads to enrich for the T cell population of interest Magnetic bead sorting is a simple solution for applications requiring enrichment of antigen-specific T

Identification of apoB‐100 Peptide‐Specific CD8+ T Cells

Pentamer staining was performed on 210 6 red blood cell–lysed mouse splenocytes using a 2‐layer approach. Biotinylated pentamers were added to the cells and incubated for 10 minutes in 25C. A control mouse H2K b SIINFEKL pentamer (ProImmune) was used as staining control. Cells were washed twice and stained with CD8a (clone KT15) and

Magnetic-activated cell sorting (PDF download), or MACS, is a procedure developed by Miltenyi Biotec to separate cells from complex mixtures using antibody-coated magnetic nanoparticl The antibodies are specific for certain cell surface markers, either expressed on your population of interest (positive selection), or expressed on undesired cell types (negative selection)

Recently improved reagents, that is, pentameric HLA class I allele/antigenic peptide complexes were developed and are commercially accessible (Pro5 MHC Class I Pentamers, Proimmune). Due to their planar configuration, all five HLA-peptide complexes, assembled through a coiled-coil domain, are available for binding to complementary TCRs, while

T cells have the capacity to eliminate tumors but the signaling pathways by which they do so are incompletely understood. T cell priming requires activation of the transcription factors AP-1, NFAT and NF-κB downstream of the TCR, but whether activation of T cell-NF-κB in vivo is required for tumor control has not been addressed. In humans and mice with progressively growing tumors, the

These pro-inflammatory NK cells are present in the inflamed lamina propria where they are enriched for integrin CD103 expression. Thus, CD161 expression identifies NK cells that may contribute to inflammatory disease pathogenesis and correlates with an innate responsiveness to cytokines in both T and NK cells. Gordon CL, Lee LN, Swadling L, Hutchings C, Zinser M, Highton AJ, Capone S, Folgori


eBioscience

Gentle, tube-based magnetic separation with our MagniSort beads enables the high-yield isolation of pure, viable and functional cells. When absolute purity is not necessary, as is often the case with in vitro stimulation of T cells, magnetic cell separation can deliver highly enriched cells faster, with significant cost-savings, and without exposure to harsh separation protocols like flow

31.08.2009Cobbold and colleagues selected CMV-specific CD8 + T cells from the blood of stem cell transplant donors using human leukocyte antigen (HLA)–peptide tetramers followed by selection with magnetic beads, and saw impressive clinical responses with eight of nine treated patients clearing their infection following infusion of tiny numbers of selected cells (median 8.6 10 3 /kg). 16 Selection

High‐gradient magnetic cell separation columns are needed for magnetization of labeled cells in a magnetic field generated by a strong external magnet. These columns are filled with a matrix of ferromagnetic steel‐wool or iron‐spheres, which focus the magnetic field lines towards their surface and thus induce strong magnetic field gradients (4 Tesla), which attract even slightly magnetic

cell separation using pro5 pentamers and magnetic beads proimmune. Epstein–Barr virus microRNAs reduce immune surveillance by . At 10 and 20 d after the first stimulation, T cells were stained with unlabeled HLA/peptide pentamers (Proimmune) for 20 min at 37 C. Counterstaining was done with CD8 and CD3specific antibodies and Pro5 fluorotag

Protocols for Cell Separation using Magnetic Beads. Guide to Cell Separation using Magnetic Beads. Column-based bead isolation for fluorescent Pentamers. Column-based bead isolation for biotin-labeled Pentamers. Tube-based bead isolation for biotin-labeled Pentamers. CD137 isolation and staining protocol . Cell Preparation Protocols

If possible, stem cell donors for transplantation are selected on the basis of their CMV serostatus. However, the cytomegalovirus-specific immune status can be further characterized by measuring CMV phosphoprotein 65–specific CD8 + T cell frequencies using tetramers, pentamers, and streptamers. We therefore investigated the specificity and

CD4+ and CD8+ T cells reside in the human bone marrow (BM) and show a heightened activation state. However, only small sample sizes are available from sources such as the iliac crest. Larger samples can be obtained from the femur in the course of hip replacement surgery. It was therefore the goal of the present study to compare the phenotype and function of BM T cells from different sources

Proimmune Co-staining of Pentamers and Intracellular Cytokines. Human Pentamer and intracellular cytokines. Murine Pentamer and intracellular cytokines . CD1d Tetramer Staining Protocols. CD1d Tetramer staining protocol . Protocols for Cell Separation using Magnetic Beads. Guide to Cell Separation using Magnetic Beads. Column-based bead isolation for

Establishment of the reversible peptide

Several pMHC multimer techniques have been developed using dimers (19, 20), tetramers (17, 21), pentamers (13, 22–24), dextramers, octamers, streptamers (8, 27) and clinimers (28, 29) for visualization, characterization and sorting of antigen-specific T cells. All of these pMHC multimers use the natural T-cell receptor (TCR) ligand (the

CD4 + T cells were isolated from spleen using magnetic bead separation (Miltenyi Biotech, Bergisch Gladbach, Germany) according to manufacturer's instructions. OT-I cells were isolated from spleen after lysing of the erythrocytes and made into a single-cell suspension by application through a 70-μm cell strainer (BD Falcon, Erembodegem, Belgium). As the OT-I mice were on a Rag1

01.01.1984An especially interesting variation of the tech- nique is the use of lectin-coated magnetic microspheres, which permit separation of the bound cells in a magnetic field.€ The application of affinity for lectins to separation of cells by phase partition has been noted above (p. 56). The technique is selective rather than specific; most studies reported to date have been concerned with the

ProImmune Ltd Original Assignee ProImmune Ltd Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.) 2002-08-21 Filing date 2003-08-14 Publication date 2015-11-24 2002-08-21 Priority to GB0219459.5 priority

Using positive selection with HLA multimers and magnetic beads, we isolated CTLs from both frozen donor material as well as third-party donors within hours. Results. At 90 days after CTL infusions 7 out of 8 patients were still living. CTLs infused from third-party donors were detected in 5 of 6 patients up to 76 days after infusion. No graft-versus-host disease associated with CTL infusions

Magnetic Bead Cell Separation ProImmune Mastering . Cell Separation using Pro5 Pentamers and Magnetic Beads In addition to detecting antigen specific T cells Pro5 MHC Class I Pentamers can be used in conjunction with magnetic beads to enrich for the T cell population of interest Magnetic bead sorting is a simple solution for applications requiring enrichment of antigen specific T cells

He received his M.Sc. in 2010 after conducting his master thesis on cell handling in microfluidic systems in the laboratory of Professor Elisabeth M. J. Verpoorte. He is currently pursuing his Ph.D. at the Stratingh Institute for Chemistry in Groningen under the supervision of Professor Ben L. Feringa. His recent research focuses on the use of molecular photoswitches in biological relevant

SpyCatcher magnetic bead preparation. For a single SpyCLIP experiment, 1 mg of Epoxy beads (1 μm diameter, BioMag, Wuxi, Jiangsu, China) was washed three times with 1 ml of PBS and re-suspended in 50 μl PBS containing 0.1 mg SpyCatcher proteins. After overnight rotation at 25C, the beads were washed three times with 1 ml of Spybeads wash buffer (PBS containing 500 mM NaCl and 0.5%

01.07.2012Tumour antigen-specific CTLs were detected by staining T-cell cultures with PE-labelled Pro5 MHC class I pentamers and FITC-labelled anti-CD8, clone LT8 (both from Proimmune Limited, UK), according to the manufacturer's instructions. Flow cytometric analysis was carried out using a FACSCalibur flow cytometer and analysed using CellQuest Pro

B cells were enriched by negative selection via magnetic depletion of non-B cells (EasySep Human B Cell Enrichment Kit, StemCell Technologies). The following fluorescently labeled antibodies were used to stain the enriched B cell fraction prior to sorting by flow cytometry: anti-CD3-PEcy5 (clone UCHT1 Beckman-Coulter #A07749), anti-CD14-PEcy5 (clone RMO52 Beckman-Coulter #A07765), anti-CD16

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