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Jes5 2a5

Manufactured by BioLegend
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The JES5–2A5 is a high-performance lab equipment designed for specialized applications. It provides reliable and precise measurements for researchers and scientists. The core function of this product is to perform specific tasks required in a laboratory setting.

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Lab products found in correlation

Perm buffer, by Thermo Fisher Scientific (1 mentions) Igg2b, by BioLegend (1 mentions) Facsaria flow cytometer, by BioLegend (1 mentions) Na pyruvate, by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) L glutamine, by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Monensin, by BioLegend (1 mentions) Jes5 16e3, by BioLegend (1 mentions) Rtk2071, by BioLegend (1 mentions) Ifn β, by PBL Assay Science (1 mentions) Il 10, by BioLegend (1 mentions) Interleukin 6 (il 6), by BioLegend (1 mentions) Tnf α, by R&D Systems (1 mentions) Lactate dehydrogenase, by Promega (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Cytoflex lx, by Beckman Coulter (1 mentions)

5 protocols using jes5 2a5

1

Intracellular Cytokine Staining of Tumor-associated B Cells

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Spleen cells from mice bearing active BCL1 tumor cells and naïve mice were suspended at 1 x 106 /mL in stimulation medium prepared using RPMI-1640 (Sigma), L-glutamine (Sigma), NEAA (Gibco), Na-pyruvate (Gibco), 10% heat-inactivated FBS β-mercaptoethanol (50 μM; Sigma), LPS (10 μg/mL; Sigma), PMA (Sigma), Ionomycin (500 ng/mL; Sigma), and monensin (2 μM; BioLegend) and incubated for 5 hours at 37°C, 5% CO2. Samples were washed with cold PBS+10%FBS and stained with antibodies against BCL1-Id, CD3 (145-2C11), CD11b (M1/70), Ter119 (TER119) [for tumor cells] and anti-CD19 (6D5) [normal B cells]. Cells were fixed and permeabilized (Perm Buffer; eBioscience) then stained with anti-IL-10 antibody [JES5-2A5] or isotype control (IgG2b; BioLegend) and analyzed on a FACSAria flow cytometer.
BitMansour A., Pop L.M, & Vitetta E.S. (2016). The Role of Regulatory B Cell-Like Malignant Cells and Treg Cells in the Mouse Model of BCL1 Tumor Dormancy. PLoS ONE, 11(12), e0167618.
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2

Antigen-Specific T Cell Responses in NOD Mice

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NOD mice at 15 weeks of age received proinsulin/alum, GABA (20 mg/mL), or combined therapy, as described above. Control groups included untreated and alum (alone)–injected mice. Ten days after the second vaccination, the frequency of antigen-specific interferon-γ (IFN-γ)–, interleukin (IL)-4–, and IL-10–secreting splenic T cells in the different groups of mice was determined by ELISPOT assays, as previously described (23 (link)), with the addition of using JES5–16E3 and JES5–2A5 (BioLegend, San Diego, CA) as capture and detection antibodies for IL-10. The tested antigens included control self-antigen mouse serum albumin (MSA), and β-cell autoantigens GAD65 (Diamyd Medical), HSPp277, and proinsulin (all at 100 µg/mL). The cells in medium alone were used as the negative controls, while cells challenged with 1 µg/mL anti-CD3 provided positive controls.
Tian J., Dang H., Nguyen A.V., Chen Z, & Kaufman D.L. (2014). Combined Therapy With GABA and Proinsulin/Alum Acts Synergistically to Restore Long-term Normoglycemia by Modulating T-Cell Autoimmunity and Promoting β-Cell Replication in Newly Diabetic NOD Mice. Diabetes, 63(9), 3128-3134.
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3

Modulating BMDM Cytokine Responses

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BMDMs were stimulated with 1 μg/ml LPS for 4 hours. In certain experiments, BMDMs were concomitantly treated with 10 μg/ml of either an IL-10 neutralizing Ab (JES5–2A5, BioLegend) or a rat IgG1, κ isotype control Ab (RTK2071, BioLegend). Cytokines were measured from BMDM culture supernatant using the following commercial kits: TNF-α (R&D), IL-6 (BioLegend), IL-10 (BioLegend), and IFN-β (PBL Assay Science).
Vural A., Nabar N.R., Hwang I.Y., Sohn S., Park C., Karlsson M.C., Blumer J.B, & Kehrl J.H. (2019). Gαi2 signaling regulates inflammasome priming and cytokine production by biasing macrophage phenotype determination. Journal of immunology (Baltimore, Md. : 1950), 202(5), 1510-1520.
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4

Antitumor CAR-T Cell Assay

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Using a construct provided by Sorrento and TNK Therapeutics, CAR-T cells were generated from mouse splenocytes as described previously44 (link) and MC38CEA cells were used as target tumour cells. Myeloid cells were isolated from LM tumour-bearing mice livers. CAR-T cells were carboxyfluorescein diacetate succinimidyl ester (CFSE, Life Technologies) labelled as per the manufacturer’s protocol. MC38CEA cells were plated at 2×104 cells/well in 96-well cell culture plate with myeloid cells and CAR-T cells in a 1:1:1 ratio as controls. Cells were treated with 100, 200 or 400 ng/mL of anti-IL-10 antibody (JES5-2A5, BioLegend). Supernatant was collected and analysed for lactate dehydrogenase as per manufacturer’s protocol (Promega). Antibodies used for flow cytometry are available in online supplemental table S4. Cells were collected for analysis using Cytoflex LX (Beckman Coulter), and postacquisition analysis was performed using FlowJo software.
Sullivan K.M., Jiang X., Guha P., Lausted C., Carter J.A., Hsu C., Labadie K.P., Kohli K., Kenerson H.L., Daniel S.K., Yan X., Meng C., Abbasi A., Chan M., Seo Y.D., Park J.O., Crispe I.N., Yeung R.S., Kim T.S., Gujral T.S., Tian Q., Katz S.C, & Pillarisetty V.G. (2022). Blockade of interleukin 10 potentiates antitumour immune function in human colorectal cancer liver metastases. Gut, 72(2), 325-337.
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5

Pneumococcal Infection in Mice

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Mice were treated with 0.1 mg/mouse of IL-10 blocking antibody (JES52A5, Biolegend, San Diego, CA, USA) or the isotype control (Rat IgG1 K Isotype Control, Biolegend, San Diego, CA, USA) through i.p. injection. After 2 h, mice were then challenged i.t. with 5 × 105 CFU of S. pneumoniae. The lungs and blood were collected 2 d postinfection to determine bacterial burden.
Siwapornchai N., Lee J.N., Tchalla E.Y., Bhalla M., Yeoh J.H., Roggensack S.E., Leong J.M, & Ghanem E.N. (2020). Extracellular adenosine enhances the ability of PMNs to kill Streptococcus pneumoniae by inhibiting IL-10 production. Journal of leukocyte biology, 108(3), 867-882.
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