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Human complement c3 elisa kit

Manufactured by Abcam
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The Human Complement C3 ELISA Kit is a quantitative in vitro diagnostic assay used to measure the levels of Complement C3 protein in human serum or plasma samples. The kit utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the target analyte.

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ELISAs (10 citations)

16 protocols using human complement c3 elisa kit

1

Quantification of Complement C3 by ELISA

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Complement C3 was quantified by a Complement C3 Human ELISA kit (cat #ab108823; abcam, Cambridge, MA, USA). Briefly, 96-well plates were incubated with C3 standard or sample for two hours at RT. After washing, wells were subsequently incubated with biotinylated C3 antibody for 1 h at RT. Following washing, peroxidase-labelled streptavidin conjugate was added to each well and incubated for 30 min. After washing, chromogen substrate was added and incubated for 10 min. After the reaction had been stopped with a stop solution, the amount of reacted substrate was measured at OD 450 nm. A standard curve was made using serial dilutions of the standard. The amount of C3 in samples was determined from the standard curve. Values were expressed as ng/mL.
Pillai A., Bruno D., Nierenberg J., Pandya C., Feng T., Reichert C., Ramos-Cejudo J., Osorio R., Zetterberg H., Blennow K, & Pomara N. (2019). Complement component 3 levels in the cerebrospinal fluid of cognitively intact elderly individuals with major depressive disorder. Biomarkers in neuropsychiatry, 1, 100007.
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2

Serum Protein Quantification in TB Patients

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The albumin human ELISA kit (Abcam, Cambridge, MA, USA; SwissProt: P02768), the lipoprotein A (APOA, LPA) human ELISA kit (Abcam, Cambridge, MA, USA; SwissProt: P08519), the human rho GDP-dissociation inhibitor 2 (ARHGDIB) ELISA kit (Cusabio Biotech. Co., LTD, China; SwissProt: P52566), the complement C3 human ELISA kit (Abcam, Cambridge, MA, USA; SwissProt: P01024), and the human Ficolin-2 (FCN2) ELISA kit (Cusabio Biotech. Co., LTD, China; SwissProt: Q15485) were used to detect protein levels in the serum. The protein concentration of 57 untreated TB patients, 53 2-month treated TB patients, 59 cured TB patients and 60 healthy controls were measured according to the manufacturer’s instructions. Serum samples were diluted with dilution factors of 1:10,000, 1:800, 1:4,000, and 1:20,000 for ALB, C3, FCN2, and LPA, respectively. ELISAs were performed according to the instructions of each kit.
Wang C., Wei L.L., Shi L.Y., Pan Z.F., Yu X.M., Li T.Y., Liu C.M., Ping Z.P., Jiang T.T., Chen Z.L., Mao L.G., Li Z.J, & Li J.C. (2015). Screening and identification of five serum proteins as novel potential biomarkers for cured pulmonary tuberculosis. Scientific Reports, 5, 15615.
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3

Baseline Cerebrospinal Fluid Biomarkers

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The lumbar puncture required participants to fast overnight and was performed between 0900 and 1000 hours. A total of 15 mL of clear CSF was collected in three polypropylene tubes. The tubes were immediately placed on ice for a maximum of 1 hour until samples were centrifuged at 4°C at 1500 rpm for 10 min. Aliquots of 0.25 mL were then placed into 1 mL polypropylene cryogenic vials and stored in Nunc eight-cell storage boxes (Nalge Nunc Internation, Rochester, NY, USA) at -80°C.
CSF and plasma levels of IL-6 and IL-8 were measured in a sandwich assay format using the Human Proinflammatory II 4-Plex Assay and a SECTOR instrument from Meso Scale Discovery Gaithersburg, MD. CSF AChE and BChE activities were determined using an inhouse method as previously described (Parnetti et al., 2011 (link)). CSF Abeta and tau determinations were described previously (Pomara et al., 2012 ). sTREM2 concentration was determined using an electrochemiluminescence immunoassay as previously described (Heslegrave et al., 2016 (link)). CSF C3 levels were measured using a Complement C3 Human ELISA kit (cat #:ab108823; abcam) as previously described (Pillai et al., 2019 (link)) and values are expressed as ng/ml. The focus of this analysis will be on the baseline lumbar puncture determination.
Pomara N., Bruno D., Plaska C.R., Pillai A., Ramos-Cejudo J., Osorio R., Imbimbo B.P., Heslegrave A., Zetterberg H, & Blennow K. (2021). Evidence of upregulation of the cholinergic anti-inflammatory pathway in late-life depression. Journal of affective disorders, 286, 275-281.
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4

Validating Differential Protein Abundance in ADA Response

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Independent assays for the nine proteins that showed differential abundance in the R/NR comparison in response to ADA were searched. For four of the proteins (TAGLN2, SH3BGRL3, TCFL5 and isoform 2 of TPM3) no commercial assay was found, for the other five proteins an ELISA was identified and used. They were the Human ABI family, member 3 (NESH) binding protein ELISA Kit (Mybiosource, San Diego, CA) for ABI3BP; the Human Tropomyosin alpha-4 chain (TPM4) ELISA kit (CUSABIO, Wuhan, Hubei Province, PRC); the ELISA Kit for Human Cofilin 1, Non Muscle (CFL1) (Cloud-Clone, Houston, TX); the Hemopexin (HPX) Human ELISA Kit (Abcam, Cambridge, UK); and the Complement C3 Human ELISA kit (Abcam). Assays were performed in duplicate according with the manufacturer instructions except for HPX, TPM4 and CFL1 that required extended incubation times to reach sufficient sensitivity. Concentrations in NR and R patients were compared after logarithmic transformation with Student t test.
Ortea I., Roschitzki B., López-Rodríguez R., Tomero E.G., Ovalles J.G., López-Longo J., de la Torre I., González-Alvaro I., Gómez-Reino J.J, & González A. (2016). Independent Candidate Serum Protein Biomarkers of Response to Adalimumab and to Infliximab in Rheumatoid Arthritis: An Exploratory Study. PLoS ONE, 11(4), e0153140.
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5

Quantification of Complement C3 by ELISA

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Complement C3 was quantified by a Complement C3 Human ELISA kit (cat #ab108823; abcam, Cambridge, MA, USA). Briefly, 96-well plates were incubated with C3 standard or sample for two hours at RT. After washing, wells were subsequently incubated with biotinylated C3 antibody for 1 h at RT. Following washing, peroxidase-labelled streptavidin conjugate was added to each well and incubated for 30 min. After washing, chromogen substrate was added and incubated for 10 min. After the reaction had been stopped with a stop solution, the amount of reacted substrate was measured at OD 450 nm. A standard curve was made using serial dilutions of the standard. The amount of C3 in samples was determined from the standard curve. Values were expressed as ng/mL.
Pillai A., Bruno D., Nierenberg J., Pandya C., Feng T., Reichert C., Ramos-Cejudo J., Osorio R., Zetterberg H., Blennow K, & Pomara N. (2019). Complement component 3 levels in the cerebrospinal fluid of cognitively intact elderly individuals with major depressive disorder. Biomarkers in neuropsychiatry, 1, 100007.
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6

Astrocyte Cytokine-Induced C3 Secretion

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Astrocytes were plated at 2×104 cells/cm2 and treated with 3 ng/ml IL-1α (Sigma), 30ng/ml TNF (Cell Signaling Tech) and 400 ng/ml C1q (MyBioSource) for 24 hours. The media was isolated and spun down to remove debris and the levels of C3 were measured using the Human Complement C3 ELISA Kit (Abcam) as per manufacturer’s instructions.
Tchieu J., Calder E.L., Guttikonda S.R., Gutzwiller E.M., Aromolaran K.A., Steinbeck J.A., Goldstein P.A, & Studer L. (2019). NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells. Nature biotechnology, 37(3), 267-275.
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7

Validating Complement Activation Assay for Nanoparticles

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Assay conditions were developed using heparinized human whole blood, plasma, and complement protected human serum, and heparinized porcine whole blood and plasma obtained from Innovative research Inc (Novi, MI). Porcine citrated plasma and human citrated plasma were also obtained from Innovative research Inc (Novi, MI). Zymosan was obtained from Sigma Aldrich. Dulbecco’s phosphate-buffered saline (without phosphate and magnesium) was obtained from Fisher Scientific. C5a human ELISA duo kit (DY2037) was obtained from R&D systems (Minneapolis, MN). Human Complement C3 ELISA Kit (ab108823) and Guinea Pig Complement C3 ELISA Kit (ab157705) were obtained from Abcam plc (Cambridge, MA).
Nanoparticles prepared for validating the assay involved l-lactide from Polysciences Inc (Warrington, PA), heterobifunctional poly(ethylene glycol) with 5000 Da molecular weight from Laysan Biosciences (Arab, AL), and d-lactide from Purac Biomaterials (Corbion, Amsterdam, Netherlands). All solvents used were ACS grade and obtained from Fisher Scientific.
Maisha N., Coombs T, & Lavik E. (2020). Development of a Sensitive Assay to Screen Nanoparticles in vitro for Complement Activation. ACS biomaterials science & engineering, 6(9), 4903-4915.
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8

Quantifying Plasma Complement C5 and C3

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Plasma levels of C5 (including the sum of C5 and C5a) and C3 (including the sum of C3, C3a and C3b) were detected using Human Complement C5 ELISA Kit and Human Complement C3 ELISA Kit (Abcam) according to the manufacturer’s instruction.
Zhang Y., Han K., Du C., Li R., Liu J., Zeng H., Zhu L, & Li A. (2021). Carboxypeptidase B blocks ex vivo activation of the anaphylatoxin-neutrophil extracellular trap axis in neutrophils from COVID-19 patients. Critical Care, 25, 51.
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9

Quantification of Soluble Biomolecules

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The culture supernatants of each cell culture were collected and stored at −80 °C until analysis. According to the manufacturer's instructions, MMP-1, MMP-14, complement component C3, amphiregulin, and thioredoxin contents in the culture supernatants were quantified using Human MMP1 Enzyme-Linked Immunosorbent Assay (ELISA) Kit (Abcam Ltd., Cambridge, UK), Human MMP14 ELISA Kit (Abcam Ltd., Cambridge, UK), Human Complement C3 ELISA Kit (Abcam Ltd., Cambridge, UK), Quantikine® Human Amphiregulin ELISA Kit (R&D Systems, Inc., Minneapolis, MN, USA), and Human TXN/Thioredoxin/TRX ELISA Kit (LifeSpan BioSciences, Inc., Seattle, WA, USA), respectively.
Fukushima K., Itaba N., Kono Y., Okazaki S., Enokida S., Kuranobu N., Murakami J., Enokida M., Nagashima H., Kanzaki S., Namba N, & Shiota G. (2021). Secreted matrix metalloproteinase-14 is a predictor for antifibrotic effect of IC-2-engineered mesenchymal stem cell sheets on liver fibrosis in mice. Regenerative Therapy, 18, 292-301.
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10

Astrocyte Cytokine-Induced C3 Secretion

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Astrocytes were plated at 2×104 cells/cm2 and treated with 3 ng/ml IL-1α (Sigma), 30ng/ml TNF (Cell Signaling Tech) and 400 ng/ml C1q (MyBioSource) for 24 hours. The media was isolated and spun down to remove debris and the levels of C3 were measured using the Human Complement C3 ELISA Kit (Abcam) as per manufacturer’s instructions.
Tchieu J., Calder E.L., Guttikonda S.R., Gutzwiller E.M., Aromolaran K.A., Steinbeck J.A., Goldstein P.A, & Studer L. (2019). NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells. Nature biotechnology, 37(3), 267-275.
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