Goat anti human fi

Manufactured by Quidel

Sourced in United States

Goat anti-human FI is a laboratory reagent used for the detection and quantification of human Ficolin-1 (FI) protein in various sample types. It is a polyclonal antibody raised in goats against human FI. This product can be used in immunoassays, Western blotting, and other applications requiring the specific recognition of human FI.

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Lab products found in correlation

Odyssey infrared system, by LI COR (2 mentions) Superblock, by Thermo Fisher Scientific (2 mentions) Nupage lds sample buffer, by Thermo Fisher Scientific (1 mentions) Lds sample buffer, by Thermo Fisher Scientific (1 mentions) Sds page, by Thermo Fisher Scientific (1 mentions)

2 protocols using goat anti human fi

Reduced FI Expression Analysis by Western Blot

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For validation of reduced recombinant FI expression levels on western blot, conditioned supernatants were denatured immediately with NuPAGE™ lithium dodecyl sulfate (LDS) Sample Buffer (ThermoFisher Scientific, USA) under non-reducing or reducing conditions (50 mm Dithiothreitol). Some supernatants were first precipitated by incubating with trichloroacetic acid (TCA) for 10 min at 4°C and then washed twice with ice-cold acetone. The pellet was dried and resuspended in 1× NuPAGE™ LDS sample buffer (ThermoFisher Scientific) prior to further use. All samples were separated on SDS-PAGE (Invitrogen, USA) and transferred to a polyvinylidene difluoride membrane. After transfer, the membrane was blocked with superblock (ThermoFisher Scientific, USA) and then incubated with goat anti-human FI (Quidel, USA). Donkey anti-goat IRDye800 (Rocklands, USA) antibody was used as detection antibody. For imaging the Odyssey infrared system was used (Li-Cor, USA).

de Jong S., Volokhina E.B., de Breuk A., Nilsson S.C., de Jong E.K., van der Kar N.C., Bakker B., Hoyng C.B., van den Heuvel L.P., Blom A.M, & den Hollander A.I. (2020). Effect of rare coding variants in the CFI gene on Factor I expression levels. Human Molecular Genetics, 29(14), 2313-2324.

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Characterizing Complement Factor I Variants

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To determine the processing of the variants located close to the FI linker region, conditioned supernatants of HEK293T cells transfected with CFI were denatured under reducing conditions with NuPAGE LDS sample buffer supplemented with 50mM Dithiothreitol. After separation on NuPAGE 4-12% Bis-Tris gels, protein was transferred to a polyvinylidene difluoride membrane. After transfer, the membrane was treated with superblock (ThermoFisher Scientific, USA) and then incubated with 500x diluted goat anti-human FI (Quidel, USA). For detection, the membrane was incubated with Donkey anti-goat IRDye800 (Rocklands, USA) antibody diluted 10,000x. For imaging the Odyssey infrared system was used (Li-Cor, USA).

de Jong S., de Breuk A., Bakker B., Katti S., Hoyng C.B., Nilsson S.C., Blom A.M., van den Heuvel L.P., den Hollander A.I, & Volokhina E.B. (2022). Functional Analysis of Variants in Complement Factor I Identified in Age-Related Macular Degeneration and Atypical Hemolytic Uremic Syndrome. Frontiers in Immunology, 12, 789897.

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