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Sample activation kit

Manufactured by R&D Systems
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The Sample Activation Kit is a laboratory tool designed to facilitate the activation of samples for various analytical procedures. It provides the necessary components to prepare samples for downstream processing and analysis. The kit includes materials and instructions to ensure consistent and reliable sample preparation, enabling researchers to obtain accurate and reproducible results.

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Lab products found in correlation

Nhp cross reactive multiplex kit, by Merck Group (2 mentions) Elisas for il 10, by Mabtech (2 mentions) Tgf β1, by R&D Systems (2 mentions) Substrate reagent pack, by R&D Systems (1 mentions) Human tgf beta 1 duoset elisa, by R&D Systems (1 mentions) Dla50, by R&D Systems (1 mentions) Db100c, by R&D Systems (1 mentions)

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Sample Activation Kit 1 (12 citations)

4 protocols using sample activation kit

1

Rhesus Plasma Cytokine Profiling

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Rhesus plasma samples were evaluated for cytokine production using a custom NHP cross-reactive multiplex kit (EMD Millipore). ELISAs for IL-10 (Mabtech) and TGFβ−1 (R&D systems) were performed according to manufacturer’s protocol. Samples were activated with 1N HCL and neutralized with 1.2N NaOH/0.5M HEPES using Sample Activation Kit (R&D systems) prior to TGFβ−1 ELISA, activated R10 levels were subtracted from all samples.
Thompson E.A., Darrah P.A., Foulds K.E., Hoffer E., Caffrey-Carr A., Norenstedt S., Perbeck L., Seder R.A., Kedl R.M, & Loré K. (2019). Monocytes Acquire the Ability to Prime Tissue-Resident T Cells via IL-10-Mediated TGF-β Release. Cell reports, 28(5), 1127-1135.e4.
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2

Quantitative Analysis of Growth Factors in 3D Hydrogel Cultures

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Medium was collected from hydrogel cell constructs on days 3, 6, 9, and 12 of culture and stored at − 80 °C until analysis. Total TGF-β1 levels were assessed using Human TGF-beta 1 DuoSet ELISA (DY240; R&D Systems, Minneapolis, MN). Similarly, GDF-15 expression was analyzed using Human GDF-15 DuoSet ELISA (DY957; R&D Systems). For the activation of latent TGF-β1, the medium was treated with Sample Activation Kit (DY010; R&D Systems). TGF-β1 and GDF-15 ELISA were performed using Substrate Reagent Pack (DY999; R&D Systems), and absorbance measurements were made at 450 nm with a 570 nm correction using a SpectraMax i3x Multi-Mode Microplate Reader. α-Amylase expression was quantified using the Human Salivary Amylase Alpha ELISA Kit (NBP2-68203; Novus Biologicals, Littleton, CO), and luminesce measurements were made using a SpectraMax i3x Multi-Mode Microplate Reader. All assays were performed in accordance with the manufacturers’ protocols. Protein levels were normalized to the number of cells in each hydrogel at each time point using the procedure detailed in the 3D Proliferation method conducted on days 1, 3, 6, 9, and 12 of culture.
Fowler E.W., van Venrooy E.J., Witt R.L, & Jia X. (2022). A TGFβR inhibitor represses keratin-7 expression in 3D cultures of human salivary gland progenitor cells. Scientific Reports, 12, 15008.
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3

Rhesus Plasma Cytokine Profiling

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Rhesus plasma samples were evaluated for cytokine production using a custom NHP cross-reactive multiplex kit (EMD Millipore). ELISAs for IL-10 (Mabtech) and TGFβ−1 (R&D systems) were performed according to manufacturer’s protocol. Samples were activated with 1N HCL and neutralized with 1.2N NaOH/0.5M HEPES using Sample Activation Kit (R&D systems) prior to TGFβ−1 ELISA, activated R10 levels were subtracted from all samples.
Thompson E.A., Darrah P.A., Foulds K.E., Hoffer E., Caffrey-Carr A., Norenstedt S., Perbeck L., Seder R.A., Kedl R.M, & Loré K. (2019). Monocytes Acquire the Ability to Prime Tissue-Resident T Cells via IL-10-Mediated TGF-β Release. Cell reports, 28(5), 1127-1135.e4.
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4

Quantification of Cytokines in HCLE Cell Conditioned Media

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To quantify cytokines in CM obtained from HCLE cells, two different approaches were taken. First, we performed ELISA assays for TGFβ1, IL-1α, and IL-1RA on CM obtained from three replicate sets of control and CM from MMC treated cells grown in Low GF HCLE media. Kits were obtained from R&D Systems (human TGFβ1: DB100C; human IL-1α/IL-1F1: DLA50, human IL1RA/IL-1F3: DRA008) and assays performed as per the manufacter’s instructions. For the ELISA assay of TGFβ1, activation was performed by treating each replicate CM using R&D Systems Sample Activation kit (DY010). ELISA samples were read using a Tecan proM200 plate reader equipped with Magellen software.
We also performed Luminex assays on HCLE-CMC and CMM and IMDM HCLE-CMC and CMM. A 96 well Invitrogen/ProcartaPlex multiplex immunoassay plate was custom ordered from ThermoFisher to allow us to simultaneously detect the following human cytokines: IL-6, IL-1α, IL-1β, TNFα, IL-17F, IL-12p70, IL-1RA, and IL-23. Cytokine concentrations in CM samples were read using a Luminex Instrument (xPONENT 3.1, Luminex 100 IS Version 2.3, Austin, TX) equipped with Millipex analyst software version 5.1 (Millipore, Burlington, MA).
Pal-Ghosh S., Karpinski B.A., Majumdar H.D., Ghosh T., Thomasian J., Brooks S.R., Sawaya A.P., Morasso M.I., Scholand K.K., de Paiva C.S., Galletti J.G, & Stepp M.A. (2022). Molecular mechanisms regulating wound repair: evidence for paracrine signaling from corneal epithelial cells to fibroblasts and immune cells following transient epithelial cell treatment with Mitomycin C. Experimental eye research, 227, 109353.
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