Ni coated plates

Manufactured by Thermo Fisher Scientific

Ni-coated plates are a type of laboratory equipment used for various applications. They feature a nickel coating on the surface, which provides specific functional properties. The core function of these plates is to serve as a versatile platform for conducting experiments and analyses. The Ni-coating can enhance certain characteristics of the plates, but a detailed description of their intended use is not available in this response.

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Lab products found in correlation

Synergy h1 microplate reader, by Agilent Technologies (1 mentions) Tmb elisa substrate, by Abcam (1 mentions)

2 protocols using ni coated plates

Influenza Vaccine IgG Antibody Titer Quantification

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Serum IgG antibody titers for the influenza vaccine were measure using an ELISA. Ni‐coated plates (Thermofisher) were coated with HA (Sino Biological) at 2.5 μg/ml in PBS for 1 h at 25°C and blocked with PBS containing 1% BSA for 1 h at 25°C. First, serum was diluted 1:250 and then four‐fold serial dilutions were performed up to 1:4,096,000 dilution. Titrations were added to plates and after 2 h at 25°C, goat–anti‐mouse IgG Fc‐HRP (1:10,000, Invitrogen, A16084) was added for 1 h at 25°C. Plates were developed with TMB substrate (TMB ELISA Substrate [High Sensitivity], Abcam). The reaction was stopped with 1 M HCl. The plates were analyzed using a Synergy H1 Microplate Reader (BioTek Instruments) at 450 nm. Endpoint titers were defined as the reciprocal of the highest serum dilution that gave an optical density above 0.1 and normalized to a pooled control used with each reading.

Saouaf O.M., Roth G.A., Ou B.S., Smith A.A., Yu A.C., Gale E.C., Grosskopf A.K., Picece V.C, & Appel E.A. (2021). Modulation of injectable hydrogel properties for slow co‐delivery of influenza subunit vaccine components enhance the potency of humoral immunity. Journal of Biomedical Materials Research. Part a, 109(11), 2173-2186.

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Sortilin and Prosaposin Binding Assay

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His-tagged recombinant human sortilin (R&D Systems, Catalog #3153-ST) or human prosaposin (PSAP) (Abcam, Catalog #ab167924) proteins were diluted in PBS and incubated with Ni-coated plates (Thermo Scientific, Catalog # 15442) for 2 hours at RT while shaking at 600 rpm. Plates were washed 3 times with PBST and blocked with 3% BSA in PBS. Post-PBST wash, plates were incubated with PTV:PGRN or Fc:PGRN proteins with serial dilution from 1000 nM to 0.03 nM in half-log steps and incubated for 2 hours at RT while shaking to attain the dose-response curve. Anti-human IgG Fc gamma HRP (Jackson Immuno Research, Catalog # 109–035-098) was used as a detection antibody and was developed with TMB substrate. 450 nm absorbance was read on the Biotek plate reader. EC50 were calculated with Graph Pad Prism 8 and curves were fitted with a 4-parameter sigmoidal curve.

Logan T., Simon M.J., Rana A., Cherf G.M., Srivastava A., Davis S.S., Yoon Low R.L., Chiu C.L., Fang M., Huang F., Bhalla A., Llapashtica C., Prorok R., Pizzo M.E., Calvert M.E., Sun E.W., Hsiao-Nakamoto J., Rajendra Y., Lexa K.W., Srivastava D.B., van Lengerich B., Wang J., Robles-Colmenares Y., Kim D.J., Duque J., Lenser M., Earr T.K., Nguyen H., Chau R., Tsogtbaatar B., Ravi R., Skuja L.L., Solanoy H., Rosen H.J., Boeve B.F., Boxer A.L., Heuer H.W., Dennis M.S., Kariolis M.S., Monroe K.M., Przybyla L., Sanchez P.E., Meisner R., Diaz D., Henne K.R., Watts R.J., Henry A.G., Gunasekaran K., Astarita G., Suh J.H., Lewcock J.W., DeVos S.L, & Di Paolo G. (2021). Rescue of a lysosomal storage disorder caused by Grn loss-of-function with a brain penetrant progranulin biologic. Cell, 184(18), 4651-4668.e25.

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