Cba flex set kit

Manufactured by BD

Sourced in United States

The CBA Flex Set kits are laboratory equipment designed for conducting clinical chemistry analyses. The kits provide the necessary components to perform biochemical assays, including reagents, consumables, and accessories. The core function of the CBA Flex Set kits is to enable the measurement and quantification of various analytes in biological samples, supporting diagnostic and research applications.

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13 protocols using cba flex set kit

Cytokine Profiling of Cell Supernatants

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The stored supernatant was thawed prior to the assay and 50 μl of the supernatant per condition was prepared according to the protocol of the BD CBA Flex Set kit to measure the concentrations of IFN-γ, IL1-β, IL-6, IL-10, VEGF and TNFα (BD Biosciences).

Ehlers F.A., Mahaweni N.M., van de Waterweg Berends A., Saya T., Bos G.M, & Wieten L. (2023). Exploring the potential of combining IL-2-activated NK cells with an anti-PDL1 monoclonal antibody to target multiple myeloma-associated macrophages. Cancer Immunology, Immunotherapy, 72(6), 1789-1801.

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Splenocyte Proliferation Assay for Malaria Vaccine Evaluation

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A splenocyte proliferation assay was performed using splenocytes removed from mice that had been immunized with rPvMSP8. The cells were stimulated with rPvMSP8 (2.5 μg/mL), Con A (5 μg/mL), LPS (10 μg/mL), or medium alone. Culture supernatants were collected after 72 h of incubation and assayed using a BD CBA Flex Set kit (BD Biosciences), according to the manufacturer’s instructions.

Cheng Y., Wang B., Changrob S., Han J.H., Sattabongkot J., Ha K.S., Chootong P., Lu F., Cao J., Nyunt M.H., Park W.S., Hong S.H., Lim C.S., Tsuboi T, & Han E.T. (2017). Naturally acquired humoral and cellular immune responses to Plasmodium vivax merozoite surface protein 8 in patients with P. vivax infection. Malaria Journal, 16, 211.

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Antigen-Presenting Cell-Mediated T-Cell Activation

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For each experiment, DC populations and naïve CD4⁺ T cells were isolated from the same donor. Sorted DCs were pelleted and resuspended in X-VIVO-15 media at 0.4∗10⁶ cells/mL and 50μL were seeded in round bottom 96-well plates. Dilutions of viral supernatants for MOI of 2 were added up to 150uL and incubated overnight at 37°C. Supernatant was then removed and previously CFSE stained autologous naïve CD4⁺ T cells were added at a ratio of 1:10 (1 DC for 10 T cells) in the presence of SuperAntigen TSST-1 (0.5 ng/mL). After 48h or 72h of co-culture, supernatants were harvested and neutralised with NP40 10X and cells were fixed in 4% PFA in PBS. Cells were then stained with Live/Dead Near-IR Red stain, CD3 AF647 and CD25 PE (BD) and analysed on a FACS Verse (BD). Supernatants were processed for IL-2 quantification using BD CBA Flex Set Kit. Data were analysed using FlowJo v10 and Prism v7 for Mac (GraphPad).

Brouiller F., Nadalin F., Bonté P.E., Ait-Mohamed O., Delaugerre C., Lelièvre J.D., Ginhoux F., Ruffin N, & Benaroch P. (2023). Single-cell RNA-seq analysis reveals dual sensing of HIV-1 in blood Axl+ dendritic cells. iScience, 26(2), 106019.

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Multiplex Cytokine Quantification

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A CBA Flex Set kit (BD Biosciences, San Jose, CA, United States) was used to assess interleukin-6 (IL-6), IL-10, interferon gamma (IFN-γ), TNF-α, and C-C motif chemokine ligand 2 (CCL2) levels in cell culture supernatant, according to the manufacturer’s instructions. Data were analyzed using the CellQuest software (BD Biosciences) and BD Pharmingen (BD Biosciences).

Cheng C., Shou Q., Lang J., Jin L., Liu X., Tang D., Yang Z, & Fu H. (2020). Gehua Jiecheng Decoction Inhibits Diethylnitrosamine-Induced Hepatocellular Carcinoma in Mice by Improving Tumor Immunosuppression Microenvironment. Frontiers in Pharmacology, 11, 809.

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Cytokine Quantification in Cell Cultures

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A CBA Flex Set kit (BD Biosciences, San Jose, CA, USA) was used to assess IL-6, IL-10, interferon gamma (IFN-γ), TNF-α, and C-C motif chemokine ligand 2 (CCL2) levels in the cell culture supernatant according to the manufacturer's instructions. Data were analyzed using CellQuest software (BD Biosciences) and BD Pharmingen (BD Biosciences).

Fu H., Tang B., Lang J., Du Y., Cao B., Jin L., Fang M., Hu Z., Cheng C., Liu X, & Shou Q. (2020). High-Fat Diet Promotes Macrophage-Mediated Hepatic Inflammation and Aggravates Diethylnitrosamine-Induced Hepatocarcinogenesis in Mice. Frontiers in Nutrition, 7, 585306.

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Cytokine and Chemokine Profiling in BALF

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Th1 (IL-2, IFN-γ)/Th2 (IL-4, IL-5, IL-13) cytokine and TNFα levels in BALF were determined by flow cytometry using CBA Flex Set kits (BD Biosciences) according to the manufacturer and as described in our previous studies [18 (link)]. Results were expressed as pg/ml BALF. Eotaxin-1(CCL11) and eotaxin-2 (CCL24) in the BALF were measured using ELISA kits (R & D Systems, Minneapolis, MN) according to the manufacturers’ recommendations.

Ge X.N., Bastan I., Ha S.G., Greenberg Y.G., Esko J.D., Rao S.P, & Sriramarao P. (2018). Regulation of eosinophil recruitment and allergic airway inflammation by heparan sulfate proteoglycan (HSPG) modifying enzymes. Experimental lung research, 44(2), 98-112.

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Quantifying Airway Cytokine Profiles

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Th1 (IL-2, IFN-γ)/Th2 (IL-4, IL-5) cytokines (including TNFα) and IL-13 were measured in the BALF by flow cytometry using CBA Flex Set kits (BD Biosciences, San Jose, CA) with a FACScan flow cytometer equipped with CellQuest Pro^TM (BD Biosciences) for data acquisition and FlowJo Software (Tree Star, Inc., Ashland, OR) for analysis or a FACSCelesta flow cytometer (for IL-13) with FACSDiva^TM Software (BD Biosciences) for data acquisition and analysis as described previously (Ge et al., 2013a (link)). Eotaxin-1 (CCL11) and eotaxin-2 (CCL24) in the BALF were measured using ELISA kits (R & D Systems, Minneapolis, MN) according to the manufacturer’s recommendations. Results were expressed as pg/ml BALF in each case.

Dileepan M., Rastle-Simpson S., Greenberg Y., Wijesinghe D.S., Kumar N.G., Yang J., Hwang S.H., Hammock B.D., Sriramarao P, & Rao S.P. (2019). Effect Of Dual sEH/COX-2 Inhibition on Allergen-Induced Airway Inflammation. Frontiers in Pharmacology, 10, 1118.

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Serum Cytokine Profiling by CBA

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Serum cytokines were measured by cytometric bead array (CBA) flex set kits (BD Biosciences, San Jose, CA): mouse tumor necrosis factor (TNF) (Cat# 558299), mouse interleukin (IL)-6 (Cat# 558301). Serum samples were diluted 1:4 using assay diluent solution provided in the kit. Capture beads and detection beads were added as described in the user guide. Cytokine concentration was measured by flow cytometry.

Khuat L.T., Vick L.V., Choi E., Dunai C., Merleev A.A., Maverakis E., Blazar B.R., Monjazeb A.M, & Murphy W.J. (2021). Mechanisms by Which Obesity Promotes Acute Graft-Versus-Host Disease in Mice. Frontiers in Immunology, 12, 752484.

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Multiplex Cytokine Analysis in Serum

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Serum cytokines were measured by CBA flex set kits (BD Biosciences, San Jose, CA, USA): mouse TNF (Cat# 558299) and mouse IL-6 (Cat# 558301). Serum samples were diluted 1:4 using the assay diluent solution provided in the kit. Capture beads and detection beads were added as described in the manufacturer’s protocol. Cytokine concentration was measured by flow cytometry with the LSRFortessa cell analyzer (BD Biosciences, San Jose, CA, USA). Mouse LBP and ST2 in the serum were measured using ELISA kits from Biometec GmbH, Greifswald, Germany (Cat# 043) and Invitrogen (by ThermoFisher Scientific, San Diego, CA, USA, Cat# 88–9334-88), respectively. Human ST2 concentrations were measured as previously described (15 (link)).

Khuat L.T., Le C.T., Pai C.C., Shields-Cutler R.R., Holtan S.G., Rashidi A., Parker S.L., Knights D., Luna J.I., Dunai C., Wang Z., Sturgill I.R., Stoffel K., Merleev A.A., More S.K., Maverakis E., Raybould H., Chen M., Canter R.J., Monjazeb A.M., Dave M., Ferrara J.L., Levine J.E., Longo D.L., Abedi M., Blazar B.R, & Murphy W.J. (2020). Obesity Induces Gut Microbiota Alterations and Augments Acute Graft-Versus-Host Disease After Allogeneic Stem Cell Transplantation. Science translational medicine, 12(571), eaay7713.

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Cytokine and Chemokine Profiling in Lung Tissue

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Th1 (IL-2, IFN-γ)/Th2 (IL-4, IL-5, IL-13) cytokine and TNF-α levels in supernatants of lung tissue lysates were determined using CBA Flex Set Kits (BD Biosciences) with a FACScan flow cytometer equipped with CellQuest Pro™ (BD Biosciences) for data acquisition and FlowJo Software (Tree Star, Inc.) for analysis. Eotaxin-1 and eotaxin-2 in the BALF and IL-33 in lung lysates was measured using ELISA kits (R&D Systems). IgE level in the BALF was measured using a mouse IgE ELISA kit (BioLegend). All kits were used as per the manufacturer’s recommendation.

Dileepan M., Ge X.N., Bastan I., Greenberg Y.G., Liang Y., Sriramarao P, & Rao S.P. (2019). Regulation of eosinophil recruitment and allergic airway inflammation by tropomyosin receptor kinase A. Journal of immunology (Baltimore, Md. : 1950), 204(3), 682-693.

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