High affinity 96 well flat bottom plates

Manufactured by Corning

High-affinity 96-well flat bottom plates are laboratory equipment designed for various applications that require high-affinity binding. These plates feature a flat bottom configuration and are typically used in experiments involving cell-based assays, protein-protein interactions, and other biochemical analyses.

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Lab products found in correlation

Wallac victor3 1420 multilabel counter, by PerkinElmer (2 mentions) Human ifnγ elisa ready set go, by Thermo Fisher Scientific (2 mentions) Multiplex bead immunoassay kit, by Thermo Fisher Scientific (1 mentions) Wallac 1420 workstation, by PerkinElmer (1 mentions) Recombinant human erbb2 her2 fc chimera protein, by R&D Systems (1 mentions)

2 protocols using high affinity 96 well flat bottom plates

PSCA-CAR T-cell Cytokine Secretion

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Varying concentrations of recombinant human PSCA protein (amino acids 23–95; Abnova) was coated overnight in 1X PBS at 4°C on high-affinity 96-well flat bottom plates (Corning). Wells were washed twice with 1X PBS, blocked with 10% FBS for 1 hr, and washed again. CAR T cells (5 × 10³ in 200 μL) were added to coated wells. Where specified, tumor targets (5 × 10³) were incubated with T cells in non-coated wells (final volume of 200 μL). Following an overnight incubation at 37°C, supernatants were harvested and processed according to the Human IFNγ ELISA Ready-SET-GO!® (eBioscience) manufacturer's protocol. Plates were read at 450 nm using the Wallac Victor3 1420 Multilabel Counter (Perkin-Elmer) and Wallac 1420 Workstation software. Alternatively, supernatants were analyzed for multiple cytokines using the Multiplex Bead Immunoassay Kit (Invitrogen) according to the manufacturer's protocol. Mouse serum was analyzed for prostate-specific antigen (PSA) using the human PSA/KLK3 ELISA (Abcam) according to the manufacturer's protocol.

Priceman S.J., Gerdts E.A., Tilakawardane D., Kennewick K.T., Murad J.P., Park A.K., Jeang B., Yamaguchi Y., Yang X., Urak R., Weng L., Chang W.C., Wright S., Pal S., Reiter R.E., Wu A.M., Brown C.E, & Forman S.J. (2017). Co-stimulatory signaling determines tumor antigen sensitivity and persistence of CAR T cells targeting PSCA+ metastatic prostate cancer. Oncoimmunology, 7(2), e1380764.

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CAR T Cell Activation Assay

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Varying concentrations of recombinant human ErbB2/HER2 Fc chimera protein (R&D Systems) were coated overnight in 1× PBS at 4°C on high-affinity 96-well flat bottom plates (Corning). Wells were washed twice with 1× PBS, blocked with 10% FBS for 1 hour, and washed again. CAR T cells (5 × 10³) were added to protein-coated wells. Where specified, tumor targets (5 × 10³) were incubated with T cells in noncoated wells (final volume of 200 μL). Following an overnight incubation at 37°C, supernatants were harvested and processed according to the Human IFNγ ELISA Ready-SET-GO! (eBioscience) manufacturer’s protocol. Plates were read at 450 nm using the Wallac Victor3 1420 Multilabel Counter (Perkin-Elmer) and Wallac 1420 Workstation software.

Priceman S.J., Tilakawardane D., Jeang B., Aguilar B., Murad J.P., Park A.K., Chang W.C., Ostberg J.R., Neman J., Jandial R., Portnow J., Forman S.J, & Brown C.E. (2017). Regional Delivery of Chimeric Antigen Receptor–Engineered T Cells Effectively Targets HER2+ Breast Cancer Metastasis to the Brain. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(1), 95-105.

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