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Hstcmag28spmx13

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HSTCMAG28SPMX13 is a type of lab equipment used for magnetic separation. It is designed to separate magnetic particles from liquid samples, which can be useful in various laboratory applications.

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Lab products found in correlation

T per, by Thermo Fisher Scientific (2 mentions) Calbiochem protease inhibitor cocktail set 1, by Merck Group (2 mentions) Prism 7, by GraphPad (2 mentions) Tissuelyser lt, by Qiagen (2 mentions)

6 protocols using hstcmag28spmx13

1

Cytokine Secretion Assay for CAR T Cells

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CAR T cells were co-cultured with target cells at a 2:1 E:T ratio in a 24-well plate. NT T cells served as controls. After 24 hr, culture supernatants were harvested, and the presence of cytokines was determined by ELISA or Multiplex assay (HSTCMAG28SPMX13; EMD Millipore, Billerica, MA, USA).
Yi Z., Prinzing B.L., Cao F., Gottschalk S, & Krenciute G. (2018). Optimizing EphA2-CAR T Cells for the Adoptive Immunotherapy of Glioma. Molecular Therapy. Methods & Clinical Development, 9, 70-80.
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2

Cytokine Secretion Assay for CAR T Cells

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CAR T cells were cocultured with target cells at a 2:1 effector-to-target (E:T) ratio in a 24-well plate. Non-transduced T cells served as controls. After 24 hours, culture supernatants were harvested, and the presence of cytokines was determined by Multiplex assay (HSTCMAG28SPMX13, EMD Millipore).
Krenciute G., Prinzing B.L., Yi Z., Wu M.F., Liu H., Dotti G., Balyasnikova I.V, & Gottschalk S. (2017). Transgenic Expression of IL15 Improves Antiglioma Activity of IL13Rα2-CAR T Cells but Results in Antigen Loss Variants. Cancer immunology research, 5(7), 571-581.
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3

Cytokine Response in CD-PBMC Preconditioning

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CD-PBMCs from 3 CD patients (#70052; STEMCELL Technologies, Vancouver, Canada) were cultured in RPMI1640 medium containing 10% exosome-depleted fetal bovine serum (A2720803; Thermo Fisher Scientific) and 1% penicillin-streptomycin. Baseline characteristics are shown in Table 7. CD-PBMCs were preconditioned with 100 μg/mL stricturing CD or non-stricturing CDSE for 2 hours and then exposed to either 0.1% TFA or 1 μg/mL elafin for 6 hours. The cells were removed by centrifugation at 10,000g for 5 minutes at 4o C. The cell supernatants were collected for a 13-plex cytokine multiplex assay (HSTCMAG28SPMX13; Millipore Sigma, St Louis, MO).
Xie Y., Fontenot L., Chupina Estrada A., Nelson B., Wang J., Shih D.Q., Ho W., Mattai S.A., Rieder F., Jensen D.D., Bunnett N.W, & Koon H.W. (2022). Elafin Reverses Intestinal Fibrosis by Inhibiting Cathepsin S-Mediated Protease-Activated Receptor 2. Cellular and Molecular Gastroenterology and Hepatology, 14(4), 841-876.
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4

Protein Expression Analysis in 3D Constructs

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3D constructs were washed with 1 mL 1X PBS and then, using a spatula, were gently scraped off of the membrane and placed in a 2 mL tube containing a 6.35 mm metal ball, T-PER (ThermoScientific, Ref. #78510), and Calbiochem protease inhibitor cocktail set I (Millipore Sigma, Cat. #539131–1VL). Samples were then homogenized using a Qiagen TissueLyser LT at 50 oscillations/second for 30 s (x2). Following 30 min incubation at 4 °C, samples were centrifuged at 12,000 RPM for 15 min at 4 °C. Supernatants were collected and stored at − 80 °C until the time of the experiment. A bead-based multiplex assay for protein detection was used to quantify expression of GM-CSF, IFN-ɣ, IL-10, IL-13, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-12 (p70), and TNF- a (MilliporeSigma, HSTCMAG28SPMX13). Data were graphed in GraphPad Prism 7 and statistics were determined using Welch’s t-test where p ≤ 0.05*, p ≤ 0.01**, p ≤ 0.001***, and p ≤ 0.0001****. Samples with values that were too low to be extrapolated from the standard curve were assigned a value of 0.01.
Brown M.E., Montgomery M.L., Kamath M.M., Nicholas S., Liu Y., Karamichos D, & Fuller K.K. (2021). A novel 3D culture model of fungal keratitis to explore host-pathogen interactions within the stromal environment. Experimental eye research, 207, 108581.
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5

Quantification of Fas-decoy and Effector Cytokines in T Cell Activation

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Transgenic Fas-decoy production was measured by Fas ELISA (R&D Systems, Minneapolis, MN, catalog no. DY326) according to manufacturer’s instructions. Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO, Houston, TX, catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FD+15 fusion molecule, samples were used in soluble Fas and IL-15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately. To measure the production of effector cytokines by T cells, supernatant harvested at 48 hrs post-activation by K562-PSCA tumor cells at an E:T of 1:1 (1×106 T cells and 1×106 tumor cells) was used to perform a 13-plex multiplex assay (Millipore Sigma, Danvers, MA, catalog no. HSTCMAG28SPMX13) according to manufacturer’s instructions.
Bajgain P., Chavez A.G., Balasubramanian K., Fleckenstein L., Lulla P., Heslop H.E., Vera J, & Leen A.M. (2022). Secreted Fas decoys enhance the antitumor activity of engineered and bystander T cells in Fas ligand–expressing solid tumors. Cancer immunology research, 10(11), 1370-1385.
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6

Protein Expression Analysis in 3D Constructs

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3D constructs were washed with 1 mL 1X PBS and then, using a spatula, were gently scraped off of the membrane and placed in a 2 mL tube containing a 6.35 mm metal ball, T-PER (ThermoScientific, Ref. #78510), and Calbiochem protease inhibitor cocktail set I (Millipore Sigma, Cat. #539131–1VL). Samples were then homogenized using a Qiagen TissueLyser LT at 50 oscillations/second for 30 s (x2). Following 30 min incubation at 4 °C, samples were centrifuged at 12,000 RPM for 15 min at 4 °C. Supernatants were collected and stored at − 80 °C until the time of the experiment. A bead-based multiplex assay for protein detection was used to quantify expression of GM-CSF, IFN-ɣ, IL-10, IL-13, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-12 (p70), and TNF- a (MilliporeSigma, HSTCMAG28SPMX13). Data were graphed in GraphPad Prism 7 and statistics were determined using Welch’s t-test where p ≤ 0.05*, p ≤ 0.01**, p ≤ 0.001***, and p ≤ 0.0001****. Samples with values that were too low to be extrapolated from the standard curve were assigned a value of 0.01.
Brown M.E., Montgomery M.L., Kamath M.M., Nicholas S., Liu Y., Karamichos D, & Fuller K.K. (2021). A novel 3D culture model of fungal keratitis to explore host-pathogen interactions within the stromal environment. Experimental eye research, 207, 108581.
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