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Recombinant human fhr 5 protein

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Recombinant human FHR-5 protein is a laboratory product produced through recombinant DNA technology. FHR-5 (Factor H-related protein 5) is a regulator of the complement system, which is part of the immune response. The recombinant protein can be used for research purposes to study the structure, function, and interactions of FHR-5.

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Lab products found in correlation

Rabbit antihuman fhr 5 antibody, by Abnova (2 mentions) Microtiter plate, by Thermo Fisher Scientific (1 mentions) H2so4, by Merck Group (1 mentions) Horseradish peroxidase conjugated rabbit anti mouse igg, by Agilent Technologies (1 mentions) Tetramethylbenzidine liquid substrate system, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions)

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Recombinant proteins

3 protocols using recombinant human fhr 5 protein

1

Quantification of FHR-5 levels by ELISA

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FHR-5 serum levels were measured with newly developed in-house ELISA method. Microtiter ELISA plates were coated with 1 μg/ml commercially available monoclonal mouse anti-human FHR-5 (IgG1, clone #390513, R&D Systems, Minneapolis, Minnesota, US) in phosphate-buffered saline (PBS) overnight, followed by blocking with PBS and 2% bovine serum albumin (BSA) the next day. Serum was diluted 1:100 in PBS containing 1% BSA and 0.05% Tween-20 and added to the plate [1 h, room temperature (RT)]. FHR-5 binding was detected using polyclonal goat anti-human FHR-5 IgG (Cat. number: AF3845, R&D Systems, Minneapolis, Minnesota, US). Concentrations of the samples were determined based on the standard curve of 2-fold dilution series of recombinant human FHR-5 protein (R&D Systems, Minneapolis, Minnesota, US). Inter-assay and intra-assay variations were determined as 11.8% and 7.8%, respectively. Specificity of the FHR-5 ELISA was confirmed by Western blot (WB) showing that the monoclonal anti-FHR-5 used as capture antibody did not detect FH or any FHR other than FHR-5 (Figure 1).
Garam N., Cserhalmi M., Prohászka Z., Szilágyi Á., Veszeli N., Szabó E., Uzonyi B., Iliás A., Aigner C., Schmidt A., Gaggl M., Sunder-Plassmann G., Bajcsi D., Brunner J., Dumfarth A., Cejka D., Flaschberger S., Flögelova H., Haris Á., Hartmann Á., Heilos A., Mueller T., Rusai K., Arbeiter K., Hofer J., Jakab D., Sinkó M., Szigeti E., Bereczki C., Janko V., Kelen K., Reusz G.S., Szabó A.J., Klenk N., Kóbor K., Kojc N., Knechtelsdorfer M., Laganovic M., Lungu A.C., Meglic A., Rus R., Kersnik Levart T., Macioniene E., Miglinas M., Pawłowska A., Stompór T., Podracka L., Rudnicki M., Mayer G., Rysava R., Reiterova J., Saraga M., Seeman T., Zieg J., Sládková E., Stajic N., Szabó T., Capitanescu A., Stancu S., Tisljar M., Galesic K., Tislér A., Vainumäe I., Windpessl M., Zaoral T., Zlatanova G., Józsi M, & Csuka D. (2021). FHR-5 Serum Levels and CFHR5 Genetic Variations in Patients With Immune Complex-Mediated Membranoproliferative Glomerulonephritis and C3-Glomerulopathy. Frontiers in Immunology, 12, 720183.
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Quantification of Plasma FHR-5 Levels

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Plasma FHR-5 was detected by enzyme-linked immunosorbent assay as previous described.20 (link),21 (link) Briefly, rabbit antihuman FHR-5 antibody (Abnova Corp., Taipei, Taiwan) was coated onto half the wells of a microtiter plate (Thermo Fisher Scientific, Waltham, MA) as the capture antibody. After blocking with 1% BSA, the standard control (recombinant human FHR-5 protein; R&D Systems, Minneapolis, MN), plasma samples and quality controls were added and incubated for 1 hour at room temperature. Binding of FHR-5 was detected using mouse anti–FHR-5 antibodies (Abnova Corp., Taipei, Taiwan). After the addition of horseradish peroxidase-conjugated rabbit antimouse immunoglobulin G (Dako), the reaction was developed using the peroxidase chromogenic substrate 3,3,5,5-tetramethylbenzididine liquid substrate system (Sigma Chemical Co., St. Louis, MO) and stopped with 1 mol/l H2SO4 (sulfuric acid) before the absorbance was measured at 450 and 570 nm.
Guo W.Y., Sun L.J., Dong H.R., Wang G.Q., Xu X.Y., Zhao Z.R, & Cheng H. (2020). Glomerular Complement Factor H–Related Protein 5 is Associated with Histologic Injury in Immunoglobulin A Nephropathy. Kidney International Reports, 6(2), 404-413.
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Quantifying Circulating FHR-5 Levels

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Circulating FHR-5 was detected by enzyme-linked immunosorbent assay as previously described. 17 Briefly, rabbit anti-human FHR-5 antibody (Abnova Corporation, Taipei, Taiwan) was coated onto half the wells of a microtitre plate (Nunc Immunoplate, Roskilde, Denmark). After blocking with 1% bovine serum albumin (Sigma Chemical Company, St. Louis, MO), the plasma samples were added and incubated for 1 hour at room temperature. Binding of FHR-5 was examined using mouse anti-human FHR-5 antibodies (Abnova Corporation). Following the addition of horseradish peroxidaseconjugated rabbit anti-mouse IgG (DakoCytomation, Glostrup, Denmark), the reaction was developed using the peroxidase chromogenic substrate 3,3 0 ,5,5 0 -Tetramethylbenzidine Liquid Substrate System (Sigma Chemical Company) and stopped with 1 mol/l sulfuric acid before the absorbance was measured at 450 and 570 nm. Serial dilutions of recombinant human FHR-5 protein (R&D Systems, Minneapolis, MN) were used to establish the standard curve, which was then used to calculate circulating FHR-5 levels.
Zhu L., Guo W.Y., Shi S.F., Liu L.J., Lv J.C., Medjeral-Thomas N.R., Lomax-Browne H.J., Pickering M.C, & Zhang H. (2018). Circulating complement factor H-related protein 5 levels contribute to development and progression of IgA nephropathy. Kidney international, 94(1).
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