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Homebrew kit

Manufactured by Quanterix
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Sourced in Sweden, United States

The Homebrew kit is a laboratory equipment product that enables users to create their own customized solutions for various applications. It provides the necessary components and tools to facilitate the preparation and modification of reagents or samples in a controlled environment. The core function of the Homebrew kit is to support the development and optimization of experimental protocols, without any specific interpretation or extrapolation on its intended use.

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Lab products found in correlation

Nf light elisa kit, by UmanDiagnostics (4 mentions) Nf light kit, by UmanDiagnostics (1 mentions) Simoa platform, by Quanterix (1 mentions) Elecsys platform, by Roche (1 mentions) Simoa gfap kit, by Quanterix (1 mentions) Human total tau kit, by Quanterix (1 mentions) Innotest elisa, by Fujirebio (1 mentions)

9 protocols using homebrew kit

1

Quantification of pan-IFNα using Simoa assay

Cited 1 time
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The Simoa pan-IFNα assay was developed using the Quanterix Homebrew kit according to the manufacturer’s instructions and using two autoantibodies specific for IFNα isolated and cloned from two APS1/APECED patients. The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/mL), and the 12H5 was biotinylated (biotin/antibody ratio = 30/1) and used as the detector at a concentration of 0.3ug/mL. The SBG revelation enzyme concentration was 150 pM. Recombinant IFNα17 was used as calibrator. The limit of detection was calculated by the mean value of all blank runs + 2SD after log conversion. The assay is fully described in Rodero et al. [13 (link)] and a video presenting all the steps is shown in Llibre et al [19 (link)].
Bondet V., Rodero M.P., Posseme C., Bost P., Decalf J., Haljasmägi L., Bekaddour N., Rice G.I., Upasani V., Herbeuval J.P., Reynolds J.A., Briggs T.A., Bruce I.N., Mauri C., Isenberg D., Menon M., Hunt D., Schwikowski B., Mariette X., Pol S., Rozenberg F., Cantaert T., Gottenberg J.E., Kisand K, & Duffy D. (2021). Differential levels of IFNα subtypes in autoimmunity and viral infection. Cytokine, 144, 155533.
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2

Quantification of pan-IFNα using Simoa assay

Cited 1 time
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The Simoa pan-IFNα assay was developed using the Quanterix Homebrew kit according to the manufacturer’s instructions and using two autoantibodies specific for IFNα isolated and cloned from two APS1/APECED patients. The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/mL), and the 12H5 was biotinylated (biotin/antibody ratio = 30/1) and used as the detector at a concentration of 0.3ug/mL. The SBG revelation enzyme concentration was 150 pM. Recombinant IFNα17 was used as calibrator. The limit of detection was calculated by the mean value of all blank runs + 2SD after log conversion. The assay is fully described in Rodero et al. [13 (link)] and a video presenting all the steps is shown in Llibre et al [19 (link)].
Bondet V., Rodero M.P., Posseme C., Bost P., Decalf J., Haljasmägi L., Bekaddour N., Rice G.I., Upasani V., Herbeuval J.P., Reynolds J.A., Briggs T.A., Bruce I.N., Mauri C., Isenberg D., Menon M., Hunt D., Schwikowski B., Mariette X., Pol S., Rozenberg F., Cantaert T., Gottenberg J.E., Kisand K, & Duffy D. (2021). Differential levels of IFNα subtypes in autoimmunity and viral infection. Cytokine, 144, 155533.
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3

Measuring Neurofilament Light in Blood and CSF

Cited 2 times
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Blood and CSF sampling procedures are described in the e-Methods. A sensitive sandwich method (NF-light® ELISA kit; UmanDiagnostics AB, Umeå, Sweden) was used to measure CSF NfL as previously described.11 (link),12 (link) NfL concentrations in blood were measured using the monoclonal antibodies and calibrator from the NfL assay, transferred onto the Simoa platform using a homebrew kit (Quanterix; Lexington, MA), as previously described.13 (link) Details of the assay performance are given in the e-Methods.
Hansson O., Janelidze S., Hall S., Magdalinou N., Lees A.J., Andreasson U., Norgren N., Linder J., Forsgren L., Constantinescu R., Zetterberg H, & Blennow K. (2017). Blood-based NfL: A biomarker for differential diagnosis of parkinsonian disorder. Neurology, 88(10), 930-937.
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4

Quantification of Neurofilament Light in CSF and Serum

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All measurements were performed by board-certified laboratory technicians in the Clinical Neurochemistry Laboratory at the Sahlgrenska University Hospital, i.e., by laboratory technicians who are licensed to perform clinical laboratory measurements by the National Board of Health and Welfare, a government agency in Sweden under the Ministry of Health and Social Affairs.
The concentration of NFL in CSF was measured with a sensitive sandwich ELISA method (NF-light ELISA kit; UmanDiagnostics AB, Umeå, Sweden) according to the ELISA kit instructions. The lower limit of quantification (LLoQ) of the assay was 31 ng/L. The intra-assay and interassay coefficients of variation were <10%.
The concentration of NFL in serum was determined with the NF-light assay, which was adapted for the Simoa platform with a Homebrew Kit (Quanterix Corp, Boston, MA). The LLoQ, which was determined by the blank mean signal at 610 SD, was 1.95 ng/L. All samples were measured in duplicate and were well above the LLoQ. The intra-assay and interassay coefficients of variation were <10%. The method is described in detail elsewhere.26 (link)
Novakova L., Zetterberg H., Sundström P., Axelsson M., Khademi M., Gunnarsson M., Malmeström C., Svenningsson A., Olsson T., Piehl F., Blennow K, & Lycke J. (2017). Monitoring disease activity in multiple sclerosis using serum neurofilament light protein. Neurology, 89(22), 2230-2237.
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5

NfL Measurement in Plasma and CSF

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Blood samples were collected by venipuncture in ethylenediaminetetraacetic acid (EDTA) tubes for plasma. After centrifugation, plasma samples were aliquoted and stored at −80°C. Plasma NfL levels were determined using the NF‐Light kit from UmanDiagnostics (UmanDiagnostics, Umeå, Sweden), transferred onto the Simoa platform using a homebrew kit (Quanterix Corp, Boston, MA). The lower limit of quantification (LLOQ), determined by the blank mean signal + 10 SD, was 1.95 pg/mL. All samples measured well above the LLOQ. The analyses were performed by a board‐certified laboratory technician using one batch of reagents with intra‐ and interassay coefficients of variation below 10% and 15%, respectively. Samples from both cohorts were analyzed using the same NfL‐Light kit batch. In the Allon cohort, lumbar punctures were performed for CSF collection and NfL measurement using an enzyme‐linked immunosorbent assay (ELISA) as described previously.7
Rojas J.C., Karydas A., Bang J., Tsai R.M., Blennow K., Liman V., Kramer J.H., Rosen H., Miller B.L., Zetterberg H, & Boxer A.L. (2016). Plasma neurofilament light chain predicts progression in progressive supranuclear palsy. Annals of Clinical and Translational Neurology, 3(3), 216-225.
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6

Plasma Neurofilament Light Quantification

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Plasma neurofilament light (NfL) data (also available for 46% of the sample; see Table 1) was analyzed by the Blennow Lab with SIMOA using a home brew kit (Quanterix Corporation) as previously described.
26 (link)
Landau S.M., Lee J., Murphy A., Ward T.J., Harrison T.M., Baker S.L., DeCarli C., Harvey D., Tosun D., Weiner M.W., Koeppe R.A, & Jagust W.J. (2024). Individuals with Alzheimer's disease and low tau burden: Characteristics and implications. Alzheimer's & Dementia, 20(3), 2113-2127.
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7

Biomarker Quantification in Plasma and Serum

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Tau was measured in plasma with the tau Simoa 2.0 assay (Quanterix) according to manufacturer’s instructions. NFL was analysed in plasma with the NFL ELISA kit from UmanDiagnostics (NF-light® ELISA kit, UmanDiagnostics AB) transferred onto the Simoa platform using a homebrew kit (Quanterix) as previously described [54 (link)]. GFAp was determined in serum using the Simoa GFAp kit (Quanterix) according to manufacturer’s instructions. The concentrations of NSE and S100B were measured in serum using immunoassays with electrochemiluminescence detection on the Elecsys platform (Roche Diagnostics) according to manufacturer’s instructions.
Pujol-Calderón F., Zetterberg H., Portelius E., Löwhagen Hendén P., Rentzos A., Karlsson J.E., Höglund K., Blennow K, & Rosengren L.E. (2021). Prediction of Outcome After Endovascular Embolectomy in Anterior Circulation Stroke Using Biomarkers. Translational Stroke Research, 13(1), 65-76.
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8

Simoa Pan-IFNα Assay Development

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The Simoa pan-IFNα assay was developed using the Quanterix Homebrew kit according to the manufacturer's instructions and using two autoantibodies specific for IFNα isolated and cloned from two APS1/APECED patients. The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3mg/mL), and the 12H5 was biotinylated (biotin/antibody ratio = 30/1) and used as the detector at a concentration of 0.3ug/mL. The SBG revelation enzyme concentration was 150pM.
Recombinant IFNα17 was used as calibrator. The limit of detection was calculated by the mean value of all blank runs + 2SD after log conversion. The assay is fully described in Rodero et al. 13 (link) and a video presenting all the steps is shown in Llibre et al 19 (link) .
Bondet V., Rodero M.P., Posseme C., Bost P., Decalf J., Haljasmägi L., Bekaddour N., Rice G., Upasani V., Herbeuval J., Reynolds J.A., Briggs T.A., Bruce I.N., Mauri C., Isenberg D., Menon M., Hunt D., Schwikowski B., Mariette X., Pol S., Rozenberg F., Cantaert T., Gottenberg J., Kisand K., & Duffy D. (2021). Differential levels of IFNα subtypes in autoimmunity and viral infection.
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9

Measurement of CSF and Plasma Tau and NF-L

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CSF levels of total tau protein were measured using the INNOTEST ELISA (Fujirebio Europe, Ghent, Belgium), while NF-L levels were measured by the NF-light® ELISA kit (UmanDiagnostics AB, Umeå, Sweden). Plasma tau was measured using the Human Total Tau kit (Quanterix, Lexington, MA) on the Simoa HD-1 analyzer (CE marker). This assay differ from the previously published Simoa tau assay (28) in that it is based on different antibodies; one monoclonal antibody for capture that reacts with a linear epitope in the mid-region of tau, and one detection antibody that reacts with a linear epitope in the N-terminal region of T-tau. NF-L concentrations in blood were measured using the same monoclonal antibodies and calibrator as in the CSF assay, but the assay was transferred onto the Simoa platform using a homebrew kit (Quanterix, Lexington, MA, USA), as previously described (9) . All CSF and plasma measurements were performed in one round of experiments using one batch of reagents by board-certified laboratory technicians who were blinded to clinical data.
Kovacs G.G., Andreasson U., Liman V., Regelsberger G., Lutz M.I., Danics K., Keller E., Zetterberg H, & Blennow K. (2017). Plasma and cerebrospinal fluid tau and neurofilament concentrations in rapidly progressive neurological syndromes: a neuropathology-based cohort. European journal of neurology, 24(11).
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