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Leukotriene b4 elisa kit

Manufactured by Cayman Chemical
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The Leukotriene B4 ELISA kit is a laboratory tool used for the quantitative determination of Leukotriene B4 levels in various sample types. It is an enzyme-linked immunosorbent assay (ELISA) designed to provide accurate and reliable measurements of this specific eicosanoid.

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Lab products found in correlation

Opti mem, by Thermo Fisher Scientific (2 mentions) Thromboxane b2 elisa kit, by Cayman Chemical (1 mentions) Prostaglandin e2 elisa kit monoclonal, by Cayman Chemical (1 mentions) Beadbug microtube homogenizer, by Benchmark Scientific (1 mentions) 3 mm zirconium beads, by Benchmark Scientific (1 mentions) Streptavidin nanobeads, by BioLegend (1 mentions) Mojosort mouse ly6g selection kit, by BioLegend (1 mentions) Mojosort mouse neutrophil isolation kit, by BioLegend (1 mentions) Anti cd40, by R&D Systems (1 mentions) 70 μm strainer, by BD (1 mentions) Recombinant mouse c5a, by R&D Systems (1 mentions) Il 10, by Thermo Fisher Scientific (1 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions) Il 1β, by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Prostaglandin e2, by Cayman Chemical (1 mentions)

6 protocols using leukotriene b4 elisa kit

1

Chondrocyte Eicosanoid Production

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We assessed eicosanoid production by chondrocytes using the Prostaglandin E2 ELISA Kit – Monoclonal, Leukotriene B4 ELISA Kit and Thromboxane B2 ELISA Kit, according to the manufacturer's recommendations (Cayman Chemical, Ann Arbor, MI, USA). The concentration of each eicosanoid was determined according to the manufacturer's recommendations.
Villas-Boas I.M., Pidde G., Lichtenstein F., Ching A.T., Junqueira-de-Azevedo I.D., DeOcesano-Pereira C., Madureira Trufen C.E., Chudzinski-Tavassi A.M., Morais K.L, & Tambourgi D.V. (2020). Human Chondrocyte Activation by Toxins From Premolis semirufa, an Amazon Rainforest Moth Caterpillar: Identifying an Osteoarthritis Signature. Frontiers in Immunology, 11, 2191.
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2

Leukotriene B4 Quantification in SH-SY5Y Cells

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After 96 hours of infection, SH-SY5Y cells supernatant was used for leukotriene B4 measurement. The experiment was made using Cayman Chemical Leukotriene B4 ELISA kit following the recommendations of manufacturer. This experiment was performed in quintuplicate, each being repeated at least three times independently.
Braz-De-Melo H.A., Pasquarelli-do-Nascimento G., Corrêa R., das Neves Almeida R., de Oliveira Santos I., Prado P.S., Picolo V., de Bem A.F., Pizato N, & Magalhães K.G. (2019). Potential neuroprotective and anti-inflammatory effects provided by omega-3 (DHA) against Zika virus infection in human SH-SY5Y cells. Scientific Reports, 9, 20119.
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3

Quantifying LTB4 in Tumor Lysates

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Tumors were dissected away from surrounding fat 24 hours post-treatment and lysed in PBS using 3 mm zirconium beads (Benchmark Scientific) in the BeadBug Microtube Homogenizer (Benchmark Scientific) for 2 cycles of 45 seconds at 3000 rpm. Crude lysate was centrifuged for 15 minutes at 16000 × g at 4°C to obtain clarified lysate. Clarified lysate was deproteinized by ethanol precipitation by adding 4 volumes of 100% ethanol, incubating on ice for 5 minutes, and centrifuging for 10 minutes at 3000 × g at 4°C. Deproteinized supernatant was transferred to a new tube, ethanol was removed by evaporation at room temperature, and samples were brought to the appropriate volume in ELISA assay buffer. LTB4 content was determined with the Leukotriene B4 ELISA kit (Cayman Chemical 520111) according to the manufacturer’s instructions and read with a Victor X4 fluorescence microplate reader (PerkinElmer).
Linde I.L., Prestwood T.R., Qiu J., Pilarowski G., Linde M.H., Zhang X., Shen L., Reticker-Flynn N.E., Chiu D.K., Sheu L.Y., Van Deursen S., Tolentino L.L., Song W.C, & Engleman E.G. (2023). Neutrophil-activating therapy for the treatment of cancer. Cancer cell, 41(2), 356-372.e10.
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4

Quantifying Ly6G+ Tumor Leukotriene B4

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B16 tumors were harvested from mice 12 hours after treatment with TNF + anti-CD40 + anti-gp75 and digested with collagenase IV and DNase I as described above. Tumor samples were then split into two halves, with one half undergoing selection with the MojoSort Mouse Ly6G Selection Kit (BioLegend) to generate Ly6G+ and Ly6G-depleted tumor samples and the other half undergoing depletion with an isotype control antibody and streptavidin nanobeads (BioLegend) to generate the “all cells” condition. These selected samples were then plated in 200μl of Opti-MEM (Gibco) and incubated for 30 minutes at 37°C. Following the incubation, the supernatant was collected, centrifuged to remove cells and debris, and analyzed for LTB4 using the Leukotriene B4 ELISA kit (Cayman Chemical 520111) as described above.
Linde I.L., Prestwood T.R., Qiu J., Pilarowski G., Linde M.H., Zhang X., Shen L., Reticker-Flynn N.E., Chiu D.K., Sheu L.Y., Van Deursen S., Tolentino L.L., Song W.C, & Engleman E.G. (2023). Neutrophil-activating therapy for the treatment of cancer. Cancer cell, 41(2), 356-372.e10.
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5

Neutrophil Activation and Leukotriene Quantification

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Bone marrow was harvested by grinding bones in a mortar and pestle and mashing through a 70 μm strainer (Falcon). Neutrophils were isolated by negative selection with the MojoSort Mouse Neutrophil Isolation Kit (BioLegend). Following isolation, neutrophils were plated in Opti-MEM (Gibco) at 1×105 cells in 100 μl and stimulated with 10 ng/ml TNF, 1 μg/ml anti-CD40, 1 μg/ml anti-gp75, and/or 50 nM recombinant mouse C5a (R&D Systems) for 30 minutes at 37°C. After 30 minutes, the supernatant was collected, centrifuged to remove cells and debris, and analyzed for LTB4 using the Leukotriene B4 ELISA kit (Cayman Chemical 520111) as described above. Additionally, stimulated neutrophils were stained for activation markers and analyzed by flow cytometry as described above.
Linde I.L., Prestwood T.R., Qiu J., Pilarowski G., Linde M.H., Zhang X., Shen L., Reticker-Flynn N.E., Chiu D.K., Sheu L.Y., Van Deursen S., Tolentino L.L., Song W.C, & Engleman E.G. (2023). Neutrophil-activating therapy for the treatment of cancer. Cancer cell, 41(2), 356-372.e10.
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6

Quantification of Inflammatory Mediators

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Cytokines TNF-α (Invitrogen, San Diego, CA), IL-1β, IL-6 and IL-10 (all from eBioscience, San Diego, CA) were quantified using sandwich enzyme-linked immunosorbent assays in accordance with the manufacturer’s instructions. To measure LTB4 and PGE2 levels, competitive assays were used (Leukotriene B4 ELISA Kit and Prostaglandin E2, both from Cayman Chemical, Ann Arbor, MI), following the manufacturer’s instructions.
Bonyek-Silva I., Nunes S., Santos R.L., Lima F.R., Lago A., Silva J., Carvalho L.P., Arruda S.M., Serezani H.C., Carvalho E.M., Brodskyn C.I, & Tavares N.M. (2020). Unbalanced production of LTB4/PGE2 driven by diabetes increases susceptibility to cutaneous leishmaniasis. Emerging Microbes & Infections, 9(1), 1275-1286.
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