Recombinant human interleukin 2

Manufactured by Roche

Sourced in Germany, Switzerland

Recombinant human interleukin-2 is a laboratory-produced cytokine that mimics the natural human interleukin-2 protein. It is a regulatory protein involved in the activation and growth of certain immune cells.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Glutamax, by Thermo Fisher Scientific (3 mentions) Ficoll paque plus, by GE Healthcare (2 mentions) Human cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Dynabeads human t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Lymphoprep density gradient medium, by STEMCELL (1 mentions) Sepmate tube, by STEMCELL (1 mentions) Dneasy blood tissue kit, by Qiagen (1 mentions) Hepes buffer, by Thermo Fisher Scientific (1 mentions) Glycerol 56 81 5, by Merck Group (1 mentions) Triton x 100 9002 93 1, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Ficoll paque premium, by BD (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Complete rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Recombinant interleukin-2 (3 citations) Human interleukin-2 (3 citations)

6 protocols using recombinant human interleukin 2

Activation and Transduction of Primary Human CD4+ T Cells

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Human primary CD4⁺ T cells were obtained from peripheral blood mononuclear cells (PBMCs) from healthy volunteer donors through the Infectious Diseases BioBank at King’s College London (ethics reference MM2-220518) under overall permission from the Southampton and South West Hampshire Research Ethics Committee (REC; B) (REC reference 19/SC/0232). PBMCs were isolated using density gradient centrifugation in SepMate tubes (STEMCELL Technologies) with Lymphoprep density gradient medium (STEMCELL Technologies). Total CD4⁺ T cells were isolated using a Human CD4⁺ T Cell Isolation Kit (Miltenyi Biotec). CD4⁺ T cells were cultured in RPMI-1640 medium with GlutaMAX and HEPES supplemented with 10% heat-inactivated autologous human serum and 1% penicillin-streptomycin (Life Technologies). Cells were then activated using Dynabeads Human T-Activator CD3/CD28 (Life Technologies) and recombinant human interleukin-2 (30 U/mL; Roche) for 48 h prior to infection.
Activated CD4⁺ T cells were transduced with lentiviral vectors (1−10 ng of p24^Gag of vector per 50,000 cells). 48 h post-transduction, the cells were fixed in 4% paraformaldehyde in DPBS before assessing transduction efficiency by flow cytometry measuring intracellular GFP expression.

Sertkaya H., Hidalgo L., Ficarelli M., Kmiec D., Signell A.W., Ali S., Parker H., Wilson H., Neil S.J., Malim M.H., Vink C.A, & Swanson C.M. (2021). Minimal impact of ZAP on lentiviral vector production and transduction efficiency. Molecular Therapy. Methods & Clinical Development, 23, 147-157.

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HTLV-1 Immortalization of Human PBMCs

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Human PBMCs were isolated from freshly collected whole blood using the Ficoll-Paque PLUS (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) density gradient medium. Two million PBMCs were cocultured with one million lethally γ-irradiated (100Gy) 729B/HTLV-1, 729B/HTLV-1p12, or 729B/HTLV-1/ΔCTCF cells in RPMI 1640 medium supplemented with 20% FBS, 10 U/mL of recombinant human interleukin-2 (Roche Diagnostics GmbH, Mannheim, Germany), glutamine, and antimicrobials in a 24-well plate. After three weeks live cells were counted and p19 in the medium was measured by ELISA every week to monitor infection and immortalization of PBMCs by HTLV-1. Immortalized PBMCs were cultured for 4-5 months and stored at -80°C. DNA was isolated using DNeasy blood tissue kit (Qiagen) for methylation, epigenetic modification and integration site studies.

Cheng X., Joseph A., Castro V., Chen-Liaw A., Skidmore Z., Ueno T., Fujisawa J.I., Rauch D.A., Challen G.A., Martinez M.P., Green P., Griffith M., Payton J.E., Edwards J.R, & Ratner L. (2021). Epigenomic regulation of human T-cell leukemia virus by chromatin-insulator CTCF. PLoS Pathogens, 17(5), e1009577.

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Virus Capture Assay Protocol

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Ultrapure grade (Type I) water was obtained through purification using
Milli-Q® Academic A-10® (Millipore). Phosphate buffered
saline (PBS) was obtained from Gibco (Grand Island, NY; 10010). Polyclonal
biotinylated goat anti-gp120 (4.0 mg/mL) for HIV-1 capture was
purchased from Abcam (Cambridge, MA; ab53937). Mouse monoclonal antibodies
reactive to proteins encoded by ORF K 8.1 A/B (0.86 mg/mL) of HHV-8
was purchased from Advanced biotechnologies (Columbia, MD; 13-212-100). Mouse
monoclonal antibodies reactive to 350/220 KDa viral glycoprotein of EBV were
purchased from Millipore (Billerica, MA; MAB10219). Mouse monoclonal antibodies
reactive to E. coli lipopolysaccharide (0.5 mg/mL) for E.
coli capture were acquired from Abcam (Cambridge, MA; AB35654). Glycerol
(56-81-5) and Triton x-100 (9002-93-1) were both purchased from Sigma (St.
Louis, MO). RPMI-1640 with L-Glutamine (10-040) was purchased from Corning
(Manassas, VA). Fetal bovine serum (100-106) was obtained from Gemini,
penicillin/streptomycin (15070-063) was purchased from Invitrogen, HEPES buffer
(15630-080) was purchased from Life Technologies (Grand Island, NY), and
recombinant human interleukin-2 (11011456001) was purchased from Roche.

Shafiee H., Kanakasabapathy M.K., Juillard F., Keser M., Sadasivam M., Yuksekkaya M., Hanhauser E., Henrich T.J., Kuritzkes D.R., Kaye K.M, & Demirci U. (2015). Printed Flexible Plastic Microchip for Viral Load Measurement through Quantitative Detection of Viruses in Plasma and Saliva. Scientific Reports, 5, 9919.

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Isolation and Activation of Primary Human and Rhesus Macaque CD4+ T Cells

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HEK293T cells, Ghost-CXCR4 (X4) cells, Jurkat T cells and HIV-1 virus were used as described previously47 (link). Whole blood samples from healthy donors were purchased from the Wuhan Blood Center (Wuhan, China). The human peripheral blood mononuclear cells (PBMCs) were separated from the whole blood by centrifugation with Ficoll-Paque Premium (BD). The primary human CD4⁺ T cells were further purified and enriched by the CD4⁺ T cell isolation Kit (Miltenyi Biotech) according to the manufacturer’s instructions and then maintained in complete RPMI medium supplemented with 10% FBS. Before transfection, the primary human CD4⁺ T cells were stimulated 2 to 3 days with anti-CD3/anti-CD28-coated plate in the presence of recombinant human interleukin-2 (20 IU/ml, Roche Applied Science). The PBMCs of whole blood from healthy Rhesus macaque (grown in the Center for Animal Experiment and ABSL-3 Laboratory, Wuhan University, China) were isolated by centrifugation with Ficoll-Paque Premium (BD). The CD4⁺ T cells were further purified with a non-human primate CD4⁺ T cell selection kit (Miltenyi Biotech). The cells were then activated with anti-CD3 (clone FN-18)/anti-CD28 (clone L293) (Invitrogen) coated on culture plate for 3 days.

Hou P., Chen S., Wang S., Yu X., Chen Y., Jiang M., Zhuang K., Ho W., Hou W., Huang J, & Guo D. (2015). Genome editing of CXCR4 by CRISPR/cas9 confers cells resistant to HIV-1 infection. Scientific Reports, 5, 15577.

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HTLV-1 Transformed T-Cell Culture Protocol

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MT-4 cells (human HTLV-1 transformed T-lymphoblasts; NIH AIDS Reagent Program) were cultured in complete RPMI 1640 medium (Thermo Fisher Scientific, Waltham, MS, USA) supplemented with 10% fetal calf serum (FCS; Merck, Darmstadt, Germany), 1% GlutaMAX, and 1% penicillin-streptomycin (both from Thermo Fisher Scientific, Waltham, MS, USA). Peripheral blood mononuclear cells (PBMC) were isolated from the blood of HIV-seronegative donors by Ficoll^® Paque Plus (GE Healthcare, Chicago, IL, USA) gradient centrifugation. PBMC were cultured in complete RPMI 1640 medium containing 100 U/mL human recombinant interleukin-2 (Hoffmann–La Roche, Basel, Switzerland).

Nifant’ev I., Siniavin A., Karamov E., Kosarev M., Kovalchuk S., Turgiev A., Nametkin S., Bagrov V., Tavtorkin A, & Ivchenko P. (2020). A New Approach to Developing Long-Acting Injectable Formulations of Anti-HIV Drugs: Poly(Ethylene Phosphoric Acid) Block Copolymers Increase the Efficiency of Tenofovir against HIV-1 in MT-4 Cells. International Journal of Molecular Sciences, 22(1), 340.

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Cell Culture Conditions for ATLL and T-Cell Leukemia Lines

Cited 1 time

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ST1, Su9T01, ATL-43Tb(−), LM-Y1(ATLL cell lines), DEL, Karpas 299, SU-DHL-1(ALK⁺ ALCL cell lines), Jurkat, and MOLT-4 (T-cell acute lymphoblastic leukemia [T-ALL] cell lines) were maintained in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. KK1, KOB, and TL-Om1 were cultured in RPMI 1640 with 10% FBS, 1% penicillin and streptomycin, and 100 IU/mL human recombinant interleukin 2 (Hoffmann-La Roche, number Ro23-6019). HEK 293T was cultured in Dulbecco modified Eagle medium with 10% FBS and 1% penicillin and streptomycin. Cell lines were engineered to express human codon-optimized S. pyogenes Cas9 using the pTO-Cas9-hygro vector (Cas9 from lentiCRISPR v2 ligated into pRCMV/TO-hygro vector) or lentiCas9-Blast. LentiCRISPR v2 and lentiCas9-Blast were gifts from Feng Zhang (Addgene, plasmid numbers 52961 and 52962, respectively). The ATLL cell lines were kindly provided by the following researchers: Michiyuki Maeda (Kyoto University; ATL43Tb(−)), Yasuaki Yamada (Nagasaki University; ST1, KK1, KOB, and LM-Y1), Tomoko Hata (Nagasaki University; ST1), Naomichi Arima (Kagoshima University; Su9T01), and Kazuo Sugamura (Tohoku University; TL-Om1). ATLL cell lines were tested for unique profiles of polymorphic DNA copy number variants.¹⁴ (link) Expression status of HBZ and Tax in ATLL cell lines were previously confirmed.¹⁴ (link)^,¹⁵ (link)

Chiba M., Shimono J., Suto K., Ishio T., Endo T., Goto H., Hasegawa H., Maeda M., Teshima T., Yang Y, & Nakagawa M. (2023). Whole-genome CRISPR screening identifies molecular mechanisms of PD-L1 expression in adult T-cell leukemia/lymphoma. Blood, 143(14), 1379-1390.

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