Mouse cxcl1 kc quantikine elisa kit

Manufactured by R&D Systems

Sourced in United States

The Mouse CXCL1/KC Quantikine ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed to measure mouse CXCL1/KC levels in cell culture supernates, cell lysates, serum, and plasma.

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11 protocols using mouse cxcl1 kc quantikine elisa kit

Quantification of Serum CXCL-1 in Mice

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After euthanasia, mice were fixed and their chest was opened using operating scissors before heart arrest. After that, about 5 mL of blood was collected using a syringe and transferred into an ice bath to prohibit clotting. Serum was isolated by centrifugation at 4500 rpm at 4 °C for 10 min. The inflammatory protein CXCL-1, also known as KC (Keratinocyte-derived chemokine), was quantified using a Mouse CXCL-1/KC Quantikine ELISA kit (R&D Systems, United States).

Kook Y.M., Hwang S., Kim H., Rhee K.J., Lee K, & Koh W.G. (2020). Cardiovascular tissue regeneration system based on multiscale scaffolds comprising double-layered hydrogels and fibers. Scientific Reports, 10, 20321.

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Quantifying TNF-α and CXCL1 in Mice

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The levels of TNF‐α and CXCL1 proteins were measured using the TNF alpha Mouse ELISA Kit (Invitrogen) and Mouse CXCL1/KC Quantikine ELISA Kit (R&D System; Minneapolis, MN, USA), respectively. Assays were performed according to manufacturers’ instructions.

Ji J., Ding K., Cheng B., Zhang X., Luo T., Huang B., Yu H., Chen Y., Xu X., Lin H., Zhou J., Wang T., Jin M., Liu A., Yan D., Liu F., Wang C., Chen J., Yan F., Wang L., Zhang J., Yan S., Wang J., Li X, & Chen G. (2024). Radiotherapy‐Induced Astrocyte Senescence Promotes an Immunosuppressive Microenvironment in Glioblastoma to Facilitate Tumor Regrowth. Advanced Science, 11(15), 2304609.

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Cytokine Quantification ELISA Protocols

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Protein concentrations were determined using the Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B) or the Mouse CXCL1/KC DuoSet (#DY453), the Mouse CXCL2/MIP-2 DuoSet (#DY452), the Mouse IL-6 DuoSet (#DY406) (R&D Systems) or the Mouse BD OptEIA IL-10 kit (BD Biosciences, #555252). The ELISAs were performed according to the manufacturer`s instructions.

Schliehe C., Flynn E.K., Vilagos B., Richson U., Swaminanthan S., Bosnjak B., Bauer L., Kandasamy R.K., Griesshammer I.M., Kosack L., Schmitz F., Litvak V., Sissons J., Lercher A., Bhattacharya A., Khamina K., Trivett A.L., Tessarollo L., Mesteri I., Hladik A., Merkler D., Kubicek S., Knapp S., Epstein M.M., Symer D.E., Aderem A, & Bergthaler A. (2014). The methyltransferase Setdb2 mediates virus-induced susceptibility to bacterial superinfection. Nature immunology, 16(1), 67-74.

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Quantification of Ear CXCL1 and CXCL2

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The amounts of CXCL1 and CXCL2 proteins in ear homogenates were analyzed by using Mouse CXCL1/KC Quantikine ELISA Kit (R&D Systems, Minneapolis, Minnesota, USA) and Mouse CXCL2 Quantikine ELISA Kit (R&D Systems), respectively, according to the manufacturer’s protocol. In brief, ear samples were homogenized for 30 sec with one 4.8-ϕ and three 3.2-ϕ beads in PBS containing protease inhibitor (P8340, Sigma) and centrifuged (10,000 rpm, 20 min, 4 °C); supernatants were collected and used for ELISA (protein concentration; 4 mg/mL). Absorbance at OD₄₅₀ and OD₅₇₀ was measured by using an iMark microplate reader (Bio-Rad, Hercules, CA, USA).

Nagatake T., Kishino S., Urano E., Murakami H., Kitamura N., Konishi K., Ohno H., Tiwari P., Morimoto S., Node E., Adachi J., Abe Y., Isoyama J., Sawane K., Honda T., Inoue A., Uwamizu A., Matsuzaka T., Miyamoto Y., Hirata S.I., Saika A., Shibata Y., Hosomi K., Matsunaga A., Shimano H., Arita M., Aoki J., Oka M., Matsutani A., Tomonaga T., Kabashima K., Miyachi M., Yasutomi Y., Ogawa J, & Kunisawa J. (2022). Intestinal microbe-dependent ω3 lipid metabolite αKetoA prevents inflammatory diseases in mice and cynomolgus macaques. Mucosal Immunology, 15(2), 289-300.

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Multiplex Biomarker Quantification

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The plasma and urine levels of mediators and biomarkers (IL-10, TNF-α, IL-6, CXCL1, CXCL2, HMGB1, IGFBP7, TIMP-2, NGAL, L-FABP, and PCX) were analyzed using commercial available ELISA kits (Mouse IL-10 Quantikine ELISA Kit, R&D Systems, M1000B; Mouse TNFα Quantikine ELISA Kit, R&D Systems, MTA00B; Mouse IL-6 Quantikine ELISA Kit, R&D Systems, M6000B; Mouse CXCL1/KC Quantikine ELISA Kit, R&D Systems, MKC00B; Mouse CXCL2/MIP2 Quantikine ELISA Kit, R&D Systems, MM200; HMGB1 ELISA, IBL-International, ST51011; IGFBP7 ELISA Kit, Antikörper-Online, ABIN816449, Mouse TIMP-2 DuoSet ELISA, R&D Systems, DY6304-05, Mouse Lipocalin-2/NGAL Quantikine ELISA Kit, R&D Systems, MLCN20, Mouse/Rat FABP1/L-FABP Quantikine ELISA Kit, R&D Systems, RFBP10: PODXL ELISA Kit Mouse, Aviva Systems Biology, OKEH03213). The ELISA plates were quantitatively analyzed on a BioTek Synergy 2 plate reader.

Thomas K., Zondler L., Ludwig N., Kardell M., Lüneburg C., Henke K., Mersmann S., Margraf A., Spieker T., Tekath T., Velic A., Holtmeier R., Hermann J., Jankowski V., Meersch M., Vestweber D., Westphal M., Roth J., Schäfers M.A., Kellum J.A., Lowell C.A., Rossaint J, & Zarbock A. (2022). Glutamine prevents acute kidney injury by modulating oxidative stress and apoptosis in tubular epithelial cells. JCI Insight, 7(21), e163161.

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Cytokine Quantification ELISA Protocols

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Quantification of CXCL1 Levels in Endothelial Cells

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The CXCL1 protein levels of primary cultured ECs extracted from Cdk5^f/f and Cdh5-Cre;Cdk5^f/f mice were detected with mouse CXCL1/KC Quantikine ELISA Kit (R&D Systems; Wang et al., 2017b (link); Taki et al., 2018 (link)). In brief, samples were incubated for 2 h and washed with Quantikine wash buffer. The absorbance was measured at 450 nm with a 540-nm correction, and the concentrations were calculated according to the manufacturer’s protocols.

Liu X.X., Yang L., Shao L.X., He Y., Wu G., Bao Y.H., Lu N.N., Gong D.M., Lu Y.P., Cui T.T., Sun N.H., Chen D.Y., Shi W.X., Fukunaga K., Chen H.S., Chen Z., Han F, & Lu Y.M. (2019). Endothelial Cdk5 deficit leads to the development of spontaneous epilepsy through CXCL1/CXCR2-mediated reactive astrogliosis. The Journal of Experimental Medicine, 217(1), e20180992.

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CXCL1 Levels in PyMT Mice

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Citrated plasma was collected from 13-week old female PyMT PAR2-WT, G37I and R38E mice. CXCL1 levels were determined with the mouse CXCL1/KC Quantikine ELISA Kit (R&D).

Graf C., Wilgenbus P., Pagel S., Pott J., Marini F., Reyda S., Kitano M., Macher-Göppinger S., Weiler H, & Ruf W. (2019). Myeloid cell-synthesized coagulation Factor X dampens anti-tumor immunity. Science immunology, 4(39), eaaw8405.

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Quantifying CXCL1 in Blood Serum and Muscle

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Blood serum CXCL1 levels were measured using a commercial CXCL1 ELISA kit (Mouse CXCL1/KC Quantikine ELISA Kit, R&D Systems) according to the manufacturer’s instructions. For quantification of muscle CXCL1, skeletal muscle was homogenized in RIPA buffer in bead-homogenizer tubes. After centrifugation, CXCL1 levels in the supernatant were quantified using the same ELISA kit.

Poller W.C., Downey J., Mooslechner A.A., Khan N., Li L., Chan C.T., McAlpine C.S., Xu C., Kahles F., He S., Janssen H., Mindur J.E., Singh S., Kiss M.G., Alonso-Herranz L., Iwamoto Y., Kohler R.H., Wong L.P., Chetal K., Russo S.J., Sadreyev R.I., Weissleder R., Nahrendorf M., Frenette P.S., Divangahi M, & Swirski F.K. (2022). Brain motor and fear circuits regulate leukocytes during acute stress. Nature, 607(7919), 578-584.

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Quantitative ELISA Assays for Chemokines

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ELISA assays were done according to manufacturer instructions by using the mouse CXCL1/KC quantikine ELISA kit (R&D, cat. no. MKC00B), the mouse CXCL4/PF4 quantikine ELISA kit (R&D, cat. no. MCX400), the mouse CXCL5/LIX quantikine ELISA kit (R&D, cat. no. MX000), and the RayBio mouse CXCL7/TCK-1 ELISA Kit (RayBiotech, cat. no. ELM-TCK1-1).

Graca F.A., Stephan A., Minden-Birkenmaier B.A., Shirinifard A., Wang Y.D., Demontis F, & Labelle M. (2023). Platelet-derived chemokines promote skeletal muscle regeneration by guiding neutrophil recruitment to injured muscles. Nature Communications, 14, 2900.

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