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Interleukin 2 (il 2)

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Sourced in Israel, United States

The IL-2 is a laboratory equipment designed for the detection and quantification of interleukin-2 (IL-2), a key cytokine involved in immune system regulation. It provides a reliable and accurate method for researchers to measure IL-2 levels in various biological samples.

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Lab products found in correlation

Il 15, by ProSpec (3 mentions) Il 21, by ProSpec (3 mentions) X vivo 10 media, by Lonza (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) X vivo 10 medium, by Lonza (2 mentions) Ef660, by Thermo Fisher Scientific (2 mentions) Human ab serum, by Valley Biomedical (2 mentions) Cd90.2 microbeads, by Miltenyi Biotec (1 mentions) Anti mouse cd3ε, by BioLegend (1 mentions) Clone 145 2c11, by BioLegend (1 mentions) Anti mouse cd28, by BioLegend (1 mentions) Clone 37, by BioLegend (1 mentions) Clone okt3, by BioLegend (1 mentions) Anti human cd3, by BioLegend (1 mentions) Anti human cd28, by BioLegend (1 mentions) Clone cd28, by BioLegend (1 mentions) Anti cd3 hit3a, by BD (1 mentions) Cd4 cd25 regulatory t cell isolation kit, by Miltenyi Biotec (1 mentions) Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)

16 protocols using interleukin 2 (il 2)

1

NK Cell Proliferation Assay

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NK cells were labeled with 2 μmol/L carboxyfluorescein succinimidyl ester (CFSE) per 1 × 10 6 cells (Invitrogen). CBCD34 + and PBNK cells were then stimulated with 200 IU IL-2 (Prospec) and CBNK cells were stimulated with 1000 IU IL-2.
Comparatively cells were also stimulated with 20 ng/mL IL-15 (Prospec) or primed with CTV-1 lysate. Proliferation was assessed on days 2, 5 and 7 post stimulation by flow cytometry.
Domogala A., Blundell M., Thrasher A., Lowdell M.W., Madrigal J.A, & Saudemont A. (2017). Natural killer cells differentiated in vitro from cord blood CD34+ cells are more advantageous for use as an immunotherapy than peripheral blood and cord blood natural killer cells. Cytotherapy, 19(6).
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2

PBMC Isolation and T-cell Expansion

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Peripheral blood mononuclear cells (PBMCs) were obtained from the whole blood of a healthy donor (according to ethics approval and written informed consent) by Ficoll (Paneco, Moscow, Russia) density centrifugation at 800× g for 30 min. The buffy coat was collected and washed twice with Dulbecco’s phosphate-buffered saline, and the cells were seeded in 10% RPMI with L-glutamine and penicillin–streptomycin (Paneco, Moscow Russia). T cells were activated with T-cell TransAct (Miltenyi, Bergisch Gladbach, Germany) according to the manufacturer’s protocol, and IL-2 (Prospec, Rehovot, Israel) was added at a concentration of 300 IU/mL. T cells were counted every two days, and a fresh medium containing IL-2 was added to maintain a cell density of approximately 1 × 106 cells/mL. After 14 days of expansion, cell populations were phenotyped by flow cytometry for CD3, CD4, and CD8 markers (Figure S1, Supplementary Materials).
Zmievskaya E.A., Mukhametshin S.A., Ganeeva I.A., Gilyazova E.M., Siraeva E.T., Kutyreva M.P., Khannanov A.A., Yuan Y, & Bulatov E.R. (2024). Artificial Extracellular Vesicles Generated from T Cells Using Different Induction Techniques. Biomedicines, 12(4), 919.
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3

Activation and Expansion of Murine T Cells

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T lymphocytes were purified from the spleens of BALB/C mice by positive selection with CD90.2 microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). T cells were activated with plate-bound anti-mouse CD3ε (2 µg/mL, clone 145-2C11) and anti-mouse CD28 (1 µg/mL, clone 37.51) antibodies (BioLegend, San Diego, CA, USA) in the presence of 100 U/mL interleukin-2 (PROSPEC, East Brunswick NJ, USA). T cells were cultured in 25 cm2 cell culture flasks with RPMI-1640 supplemented with penicillin–streptomycin and 10% foetal bovine serum medium in a total volume of 10 mL and maintained at 37 °C under 5% CO2 atmosphere. The cells were diluted up to 1 × 106 cells/mL by adding fresh culture medium supplemented with 20 U/mL interleukin-2 on day 3. On day 4, the T cells were used for co-culture with CT26 cells.
Murayama M., Hosonuma M., Kuramasu A., Kobayashi S., Sasaki A., Baba Y., Narikawa Y., Toyoda H., Isobe J., Funayama E., Tajima K., Sasaki A., Maruyama Y., Yamazaki Y., Shida M., Hamada K., Hirasawa Y., Tsurui T., Ariizumi H., Ishiguro T., Suzuki R., Ohkuma R., Kubota Y., Horiike A., Sambe T., Tsuji M., Wada S., Kobayashi S., Shimane T., Tsunoda T., Kobayashi H., Kiuchi Y, & Yoshimura K. (2024). Isobutyric acid enhances the anti-tumour effect of anti-PD-1 antibody. Scientific Reports, 14, 11325.
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4

Isolation and Activation of Primary T Cells

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PBMCs were isolated from the peripheral blood of healthy donors using Ficoll-Paque density gradient centrifugation. T cells were activated with plate-bound anti-human CD3 (2 µg/mL, clone OKT3) and anti-human CD28 (2 µg/mL, clone CD28.2) antibodies (BioLegend, San Diego, CA, USA) in the presence of 100 U/mL interleukin-2 (PROSPEC, East Brunswick NJ, USA). T cells were cultured in 25 cm2 cell culture flasks with iMediam for T medium (GC LYMPHOTEC, Tokyo, Japan) in a total volume of 10 mL and maintained at 37 °C in 5% CO2 atmosphere. The cells were diluted up to 5 × 105 T cells/mL by adding fresh culture medium supplemented with 25 U/mL interleukin-2 on days 4, 7, and 10. On day 14, the T cells were used for co-culture with T3M-1 Clone2 cells. All protocols and experiments involving primary PBMCs were approved by the Institutional Review Board of the Showa University. Informed consent was obtained from all volunteers.
Murayama M., Hosonuma M., Kuramasu A., Kobayashi S., Sasaki A., Baba Y., Narikawa Y., Toyoda H., Isobe J., Funayama E., Tajima K., Sasaki A., Maruyama Y., Yamazaki Y., Shida M., Hamada K., Hirasawa Y., Tsurui T., Ariizumi H., Ishiguro T., Suzuki R., Ohkuma R., Kubota Y., Horiike A., Sambe T., Tsuji M., Wada S., Kobayashi S., Shimane T., Tsunoda T., Kobayashi H., Kiuchi Y, & Yoshimura K. (2024). Isobutyric acid enhances the anti-tumour effect of anti-PD-1 antibody. Scientific Reports, 14, 11325.
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5

Isolation and Activation of Regulatory T Cells

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CB Tregs were isolated after ficoll using the CD4+CD25+ Regulatory T cell isolation kit (Miltenyi Biotec)49 . Purity ranged from 90 to 96%. For TCR stimulation, plates were coated for 2 h at 37 °C with 10 μg/mL anti-CD3 (HIT3a, BD Biosciences) and Tregs were cultured with 10 μg/mL anti-CD28 (CD28.2, BD Biosciences) and 1000 IU/mL IL-2 (Prospec) for 24 h at 37 °C. Activation was assessed by analyzing CD69, CTLA4, GITR and LAP expression by flow cytometry (Figure S6). Tregs were washed and added to HSC cultures at a 1:4 ratio, except from day 2 in which a 1:1 ratio was used. For transwell cultures, Tregs were placed in the upper chamber of HTC Transwell plates (Corning). The bottom chamber was coated with irradiated EL08.1D2 cells and HSC added as previously described.
Pedroza-Pacheco I., Shah D., Domogala A., Luevano M., Blundell M., Jackson N., Thrasher A., Madrigal A, & Saudemont A. (2016). Regulatory T cells inhibit CD34+ cell differentiation into NK cells by blocking their proliferation. Scientific Reports, 6, 22097.
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6

Co-encapsulation of NK92 and K562 Cells

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NK92 cells and K562 human leukemia cells were acquired from ATCC. NK-92 were cultured using X-VIVO10 media (Lonza) supplemented with 10% heat-inactivated FBS (Gibco) and 500 IU/mL IL2 (Prospec). K562 were cultured in RPMI1640 media supplemented with 10% heat-inactivated FBS and 1% antibiotic-antimycotic mixture (Gibco). Cell medias were combined at a 1:1 ratio for droplet co-encapsulations. Cells were kept at 37°C and 5% CO2. Both NK92 cells and K562 human leukaemia cells were used in a concentration of 3×106 cells/mL.
Agnihotri S.N., Ugolini G.S., Sullivan M.R., de Ganzo Y.Y., Lim J.W, & Konry T. (2022). Droplet Microfluidics for Functional Temporal Analysis and Cell Recovery on Demand using Microvalves: Application in Immunotherapies for Cancer. Lab on a chip, 22(17), 3258-3267.
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7

Glioma Patients' Immune Response Profiling

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Whole blood was obtained at the time of diagnosis from patients with gliomas (i.e., prior to surgery, radiation and chemotherapy), diluted 1:1.5 with RPMI 1640 l-glutamine (2 mM) with antibiotics (penicillin, 100 IU/mL, and streptomycin, 100 µg/mL) (Life Technologies, Carlsbad, USA) and incubated (i) without cytokines (RPMI only), (ii) with a IL-7 (10 ng/ml)/IL-2 (500 IU/ml) cytokine cocktail or (iii) with a IL-2 (1000 IU/ml)/IL-15 (10 ng/ml)/IL-21 (10 ng/ml) cytokine cocktail (Prospec, Ness Ziona, Israel). The diluted blood was co-incubated in pre-coated plates with a panel of different TAA and viral antigens (Supplementary Table 1) for 7 days at 37 °C and 5% CO2 as described previously [23 (link), 24 (link)]. To gauge the basal IFN-γ production to NY-ESO-1, survivin as well as commonly recognized target antigens, blood was incubated with medium (negative control) and non-tumor-related antigens, i.e., the Epstein–Barr virus nuclear antigen 3 (EBNA-3). Positive controls were phytohemagglutinin (PHA, Sigma-Aldrich), OKT3 (anti-CD3 monoAb, Biolegend, CA, USA, 30 ng/ml) and SEA + SEB (staphylococcal enterotoxin A and B). IFN-γ production was then quantified in the cell culture supernatant by ELISA (Mabtech, Stockholm, Sweden).
Liu Z., Poiret T., Persson O., Meng Q., Rane L., Bartek J J.r., Karbach J., Altmannsberger H.M., Illies C., Luo X., Harvey-Peredo I., Jäger E., Dodoo E, & Maeurer M. (2017). NY-ESO-1- and survivin-specific T-cell responses in the peripheral blood from patients with glioma. Cancer Immunology, Immunotherapy, 67(2), 237-246.
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8

Expansion and Maintenance of Multiple Myeloma and NK Cell Lines

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RPMI-8226 (multiple myeloma (MM) cell line) was purchased from American Type Culture Collection (ATCC, Manassas, VA) and maintained in RPMI-1640 medium supplemented with 10% Fetal Bovine Serum and 1% Antibiotic-Antimycotic solution (Corning Cellgro, Manassas, VA). NK-92 cells were obtained from Nantkwest Inc (Woburn, MA) and maintained in X-Vivo 10 media (Lonza, NJ) supplemented with 5% human serum and 500 IU/mL IL-2 (ProSpec Bio, East Brunswick, NJ). All cells were grown at 37°C under 5% CO2 in a humidified atmosphere. Cells were routinely passaged every three days and harvested at a density of 1×106 viable cells/mL.
Primary human NK cells were sorted via FACS from peripheral blood of a healthy donor by labeling PBMCs with FITC-conjugated CD56 antibody (BD Biosciences, San Jose, CA). Freshly isolated cells were maintained in RPMI-1640 media containing 10% FBS, 1% antibiotic and 100 U/mL IL-2 (Roche Diagnostics, Indianapolis, IN).
Sarkar S., McKenney S., Sabhachandani P., Adler J., Hu X., Stroopinksy D., Rosenblatt J., Avigan D, & Konry T. (2018). Anti-myeloma activity and molecular logic operation by Natural Killer cells in microfluidic droplets. Sensors and actuators. B, Chemical, 282, 580-589.
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9

Electroporation of NK-92 Cells with CD19CAR mRNA

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Example 2

NK-92 cells were grown in X-Vivo10 medium (Lonza, Basel, Switzerland) supplemented with 5% Human AB Serum (Valley Biomedical, Winchester, Va.) and 500 IU/mL IL-2 (Prospec, Rehovot, Israel). Cells were electroporated with mRNA using the Neon™ electroporation device (Life Technologies, Carlsbad, Calif.), following the manufacturer's parameters for NK-92 cells (1250 V, 10 ms, 3 pulses) and using 5 μg of mRNA per 106 cells in a volume of 100 μl. Electroporated cells were maintained in medium (same as above) for 20 hours (h).

The CD19CAR expression on the NK-92 cell surface was determined by flow cytometry using anti-scFv antibody labeled with eF660 (eBioscience, San Diego, Calif.). FIG. 2A shows the % expression of the indicated CD19CAR in the NK-92 cell population. FIG. 2B shows the median fluorescence intensity (MFI, minus background) of cells electroporated with the indicated CD19CAR. As can be taken from FIGS. 2A and 2B, CAR FcRe unexpectedly had the highest percentage of cells (75.2%) expressing CD19CAR at the cell surface, as well as the highest MFI (quantity of expressed CAR on a recombinant cell), followed by 28_3z (61.7%).

US11643452B2. Fc-epsilon car (2023-05-09). ImmunityBio, Inc. [US]. Inventors: Laurent H. Boissel [US], Hans G. Klingemann [US], Abhijit Dandapat [US], Himani Chinnapen [US].
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10

Electroporation of NK-92 Cells with CD19CAR

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Example 2

NK-92 cells were grown in X-Vivo10 medium (Lonza, Basel, Switzerland) supplemented with 5% Human AB Serum (Valley Biomedical, Winchester, Va.) and 500 IU/mL IL-2 (Prospec, Rehovot, Israel). Cells were electroporated with mRNA using the Neon™ electroporation device (Life Technologies, Carlsbad, Calif.), following the manufacturer's parameters for NK-92 cells (1250 V, 10 ms, 3 pulses) and using 5 μg of mRNA per 106 cells in a volume of 100 μl. Electroporated cells were maintained in medium (same as above) for 20 hours (h).

The CD19CAR expression on the NK-92 cell surface was determined by flow cytometry using anti-scFv antibody labeled with eF660 (eBioscience, San Diego, Calif.). FIG. 2A shows the % expression of the indicated CD19CAR in the NK-92 cell population. FIG. 2B shows the median fluorescence intensity (MFI, minus background) of cells electroporated with the indicated CD19CAR. As can be taken from FIGS. 2A and 2B, CAR FcRe unexpectedly had the highest percentage of cells (75.2%) expressing CD19CAR at the cell surface, as well as the highest MFI (quantity of expressed CAR on a recombinant cell), followed by 28_3z (61.7%).

US11547727B2. Chimeric antigen receptor-modified NK-92 cells (2023-01-10). ImmunityBio, Inc. [US]. Inventors: Laurent H. Boissel [US], Hans G. Klingemann [US].
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