Detachabead cd19

Manufactured by Thermo Fisher Scientific

Sourced in United States

DETACHaBEAD CD19 is a magnetic bead-based cell separation product designed for the isolation of CD19-positive cells from various sample types. The product utilizes magnetic beads coated with antibodies specific to the CD19 antigen, which allows for the efficient separation and enrichment of CD19-expressing cells.

Automatically generated - may contain errors

Lab products found in correlation

11 protocols using detachabead cd19

Isolation and Maintenance of Primary B Cells and EBV-Transformed Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary B cells were isolated from peripheral blood using CD19 Dynabeads and CD19 Detachabeads (Invitrogen), as previously described (27 (link)).
Cultures of mouse thymic stromal cells (OP9) expressing the human Delta-like ligand 1 (OP9-DLL1 cells) or control green fluorescent protein (GFP; OP9-GFP cells) (kindly provided by Graham Anderson, The University of Birmingham) were maintained for up to 20 passages in Opti-MEM medium (Gibco, Life Technologies) with 20% fetal calf serum (FCS).
BNHL-1 cells were derived from a bone marrow sample from a patient being treated for low-grade B cell non-Hodgkin's lymphoma (NHL). Cells of the BNHL-1 cell line were phenotypically the same as cells of the low-grade B cell NHL observed in the patient. These cells were infected with EBV and were continually maintained on the OP9-DLL1 cells, unless specified otherwise. An EBV-negative counterpart was maintained in the same manner.
293 cells containing recombinant wild-type EBV strain 2089 were maintained in RPMI, 10% FCS, and 100 μg/ml hygromycin. Akata cells containing recombinant wild-type EBV Akata were maintained in RPMI, 10% FCS, and 50 ng/ml G418.

Rowe M., Raithatha S, & Shannon-Lowe C. (2014). Counteracting Effects of Cellular Notch and Epstein-Barr Virus EBNA2: Implications for Stromal Effects on Virus-Host Interactions. Journal of Virology, 88(20), 12065-12076.

+ Open protocol

+ Expand

Isolation and Transformation of B-Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Blood samples were collected from donors and peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Hypaque centrifugation. Lymphoblastoid cell lines (LCLs) were generated by culturing PBMCs in standard LCL culture media (RPMI 1640 medium with 100 IU/ml penicillin, 100 mg/ml streptomycin, L-glutamine and 10% foetal calf serum) supplemented with 0.1ug/ml cyclosporin A (CSA; Sandimmune; Novartis Pharmaceuticals) with the addition of B95.8 strain EBV or a recombinant EBV lacking the BZLF1 gene (to generate B95.8 or BZLF1-K/O LCLs respectively). B-cells were isolated from PBMC immediately before use in experiments using anti-CD19 Dynabeads, and CD19 Detach-a-beads (Invitrogen) following the manufacturer’s instructions. B-cell purity was assessed by flow cytometry and purified B-cell preparations typically contained at least 95% B-cells.

Dowell A.C., Haigh T.A., Ryan G.B., Turner J.E., Long H.M, & Taylor G.S. (2021). Cytotoxic CD4+ T-cells specific for EBV capsid antigen BORF1 are maintained in long-term latently infected healthy donors. PLoS Pathogens, 17(12), e1010137.

+ Open protocol

+ Expand

Isolation of Peripheral Blood B-cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peripheral blood B-cells were isolated from healthy donors’ blood obtained from the Gulf Coast Blood Center (Houston, TX) by using CD19-positive magnetic beads and were released with the competitive CD19 DETACHaBEAD according to the manufacturer’s instructions (Invitrogen-Life Technologies, San Diego, CA). All procedures were performed under The University of Texas MD Anderson Cancer Center IRB-approved clinical protocol LAB08-0190.

Mathur R., Sehgal L., Braun F.K., Berkova Z., Romaguerra J., Wang M., Rodriguez M.A., Fayad L., Neelapu S.S, & Samaniego F. (2015). Targeting Wnt pathway in mantle cell lymphoma-initiating cells. Journal of Hematology & Oncology, 8, 63.

+ Open protocol

+ Expand

Cell Line Authentication and Lymphocyte Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Granta-519 cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ). Remaining cell lines were obtained from the American Type Culture Collection (ATCC). Cells were maintained in RPMI-1640 (Hyclone) with 10% fetal bovine serum (Atlanta Biologicals) (RPMI-1640/10% FBS) in 5% CO₂ atmosphere at 37°C. Cell lines were authenticated by STR analysis (MD Anderson Cancer Center Characterized Cell Line Core) and regularly tested for mycoplasma (Lonza).
Peripheral blood B-lymphocytes were isolated from healthy donors’ blood, obtained from Gulf Coast Blood Center (Houston, TX), with CD19-positive magnetic beads and released with the competitive CD19 DETACHaBEAD (Invitrogen).

Sehgal L., Mathur R., Braun F.K., Wise J.F., Berkova Z., Neelapu S., Kwak L.W, & Samaniego F. (2014). FAS-antisense 1 lncRNA and production of soluble versus membrane Fas in B-cell lymphoma. Leukemia, 28(12), 2376-2387.

+ Open protocol

+ Expand

Mantle Cell Lymphoma Patient Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mantle cell lymphoma patients’ samples and clinical information were collected and published under The University of Texas MD Anderson Cancer Center IRB-approved clinical protocol LAB08-0190 for use of human tissues, with the written informed consent of all patients. Information about the patients is shown in Additional file 1: Table S1. Normal B lymphocytes were isolated from peripheral blood of healthy donors’ blood, obtained from Gulf Coast Blood Center (Houston, TX, USA), with CD19+ magnetic beads and released with DETACHaBEAD CD19 (Invitrogen, Grand Island, NY, USA).

Wang X., Sehgal L., Jain N., Khashab T., Mathur R, & Samaniego F. (2016). LncRNA MALAT1 promotes development of mantle cell lymphoma by associating with EZH2. Journal of Translational Medicine, 14, 346.

+ Open protocol

+ Expand

Isolation and Culture of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

PBMCs were isolated from anonymous healthy donor buffy coats provided by the Oslo University Hospital Blood Bank (project number: F8). NK cells were isolated from PBMCs using CD56-reactive microbeads and an AutoMACS Pro Separator according to the manufacturer’s instructions (Miltenyi Biotec, GmbH). Purification was verified by phenotypic analysis of the surface marker CD56. B and T cells were isolated from PBMCs using the Dynabeads™ CD19 PanB with DETACHaBEAD^® CD19 and CD4-/CD8-Dynabeads™ positive isolation kits, respectively, as described by the manufacturer (Invitrogen Dynal AS, Oslo, Norway). The cells were cultured in either RPMI-1640 supplemented with 10% heat-inactivated FBS and 1% PS or in X-vivo 15 medium (Lonza, Basel, Switzerland) supplemented with 1% PS. Approval for obtaining buffy coats from healthy volunteers was granted by the Oslo University Hospital Ethics Committee.

Sioud M, & Olberg A. (2023). Antibody Surface Profiling Identifies Glycoforms in Multiple Myeloma as Targets for Immunotherapy: From Antibody Derivatives to Mimetic Peptides for Killing Tumor Cells. Cancers, 15(7), 1934.

+ Open protocol

+ Expand

Isolation and Sorting of B Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mononuclear cells were isolated from PB and SF using a density centrifugation with LSM (MP Biomedicals, LLC, Santa Ana, CA, USA). B cells were isolated by positive selection with Dynabeads M450 CD19 and DETACHaBEAD CD19 (Invitrogen, Carlsbad, CA, USA) as previously described [19 (link)]. As we previously showed, only negligible levels of artificial activation of B cells occurred immediately after positive selection [20 (link)]. To avoid unwanted further stimulation, cells were normally rested on ice prior to any stimulation. Isolated B cells exhibited greater than 99.5 % viability and more than 95 % purity, confirmed by flow cytometry. Cells were stained with mouse or rabbit monoclonal antibody (mAb) against human CD19, IgD, CD27, CD80, CD86, CD183 (CXCR3), CD254 (RANKL) and CD307d (FcRL4) (all from BioLegend, San Diego, CA, USA). Naïve (IgD⁺CD27^–), IgD⁺-memory (IgD⁺CD27⁺), double-negative (IgD^–CD27^–) and switched-memory (IgD^–CD27⁺) B cell subsets were purified by flow cytometry.

Ota Y., Niiro H., Ota S.I., Ueki N., Tsuzuki H., Nakayama T., Mishima K., Higashioka K., Jabbarzadeh-Tabrizi S., Mitoma H., Akahoshi M., Arinobu Y., Kukita A., Yamada H., Tsukamoto H, & Akashi K. (2016). Generation mechanism of RANKL+ effector memory B cells: relevance to the pathogenesis of rheumatoid arthritis. Arthritis Research & Therapy, 18, 67.

+ Open protocol

+ Expand

Isolation and Purification of CLL-B Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The CLL-B cells were collected from blood samples at the Moores-UCSD Cancer Center in compliance with the Declaration of Helsinki and after approval of the UC San Diego Institutional Review Board (IRB) [24 ].Peripheral blood mononuclear cells (PBMC) from CLL patients were isolated using Ficoll-Hypaque gradient density centrifugation (Cat# 17-1440-03, GE Healthcare Life Science). For caspase activation assays, the CLL-B cells were purified by positive selection using Dynabeads CD19 pan B (Cat# 11143D, Invitrogen) and DETACHaBEAD CD19 (Cat# 12506D, Invitrogen) according to the manufacturer’s protocol. For the other assays, fresh or frozen PBMCs were used and cells were stained with CD19/CD5 antibodies for detection of double positive CLL-B cells.

Kashyap M.K., Amaya-Chanaga C.I., Kumar D., Simmons B., Huser N., Gu Y., Hallin M., Lindquist K., Yafawi R., Choi M.Y., Amine A.A., Rassenti L.Z., Zhang C., Liu S.H., Smeal T., Fantin V.R., Kipps T.J., Pernasetti F, & Castro J.E. (2017). Targeting the CXCR4 pathway using a novel anti-CXCR4 IgG1 antibody (PF-06747143) in chronic lymphocytic leukemia. Journal of Hematology & Oncology, 10, 112.

+ Open protocol

+ Expand

Isolation of B Cells and T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peripheral blood mononuclear cells (PBMCs) were isolated from heparinized peripheral blood using Ficoll–Paque gradient centrifugation (Sigma-Aldrich, St. Louis, MO, USA). CD19⁺ B cells were isolated by magnetic bead-based positive selection using Dynabeads and DETACHaBEAD CD19 (Invitrogen). CD4⁺ T cells were isolated with Dynabeads CD4 (invitrogen) and CD25⁺ T cells were depleted from the CD4⁺ T cell population using Dynabeads CD25 (invitrogen). All isolations were done according to the manufacturer’s instructions. The purity of isolated CD19⁺ B cells was consistently >95% as analyzed by flow cytometry.

Lemarquis A.L., Einarsdottir H.K., Kristjansdottir R.N., Jonsdottir I, & Ludviksson B.R. (2018). Transitional B Cells and TLR9 Responses Are Defective in Selective IgA Deficiency. Frontiers in Immunology, 9, 909.

+ Open protocol

+ Expand

B Cell Isolation from Blood

Check if the same lab product or an alternative is used in the 5 most similar protocols

Using the same blood samples (both MCL patients and healthy controls) collected, B lymphocytes were isolated using CD19+ magnetic beads and the cells were released using DETACHaBEAD CD19 (Invitrogen). Samples with the purify and/or viability below 95% were selected for use.

Tang X., Long Y., Xu L, & Yan X. (2020). LncRNA MORT Inhibits Cancer Cell Proliferation and Promotes Apoptosis in Mantle Cell Lymphoma by Upregulating miRNA-16. Cancer Management and Research, 12, 2119-2125.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!