Human albumin elisa quantitation set

Manufactured by Fortis Life Sciences

Sourced in United States

The Human Albumin ELISA Quantitation Set is a laboratory equipment product designed for the quantitative determination of human albumin levels using the enzyme-linked immunosorbent assay (ELISA) technique. The set includes the necessary reagents and materials to perform the ELISA assay, allowing for the accurate measurement of albumin concentrations in various samples.

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37 protocols using human albumin elisa quantitation set

Measuring Organoid Secretion Levels

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To measure albumin secretion level of HLO, 200 μL of culture supernatant was collected from HLO embedded in Matrigel. For IL-6 and P3NP, 20-30 organoids were seeded and cultured on an ultra-low attachment multiwell plates 96 well plate (Corning). The supernatants were collected after 24 (for albumin), 96 (for IL-6) or 120 hours (for P3NP) from the culture and stored at −80 °C until use. To define the exact number of organoids in each well, the organoids were captured on The KEYENCE BZ-X710 Fluorescence Microscope. The supernatant was assayed with Human Albumin ELISA Quantitation Set (Bethyl Laboratories, Inc., TX, USA), Human IL-6 ELISA Kit (Biolegend, CA, USA), and Human N-terminal procollagen III propeptide, PIIINP ELISA Kit (My BioSource, CA, USA) according to the manufacturer’s instructions. Secreted level of IL-6 and P3NP was normalize the by the number of organoids in the analyzed well.

Ouchi R., Togo S., Kimura M., Shinozawa T., Koido M., Koike H., Thompson W., Karns R., Mayhew C., McGrath P.S., McCauley H.A., Zhang R.R., Lewis K., Hakozaki S., Ferguson A., Saiki N., Yoneyama Y., Takeuchi I., Mabuchi Y., Akazawa C., Yoshikawa H.Y., Wells J.M, & Takebe T. (2019). Modeling Steatohepatitis in Humans with Pluripotent Stem Cell-Derived Organoids. Cell metabolism, 30(2), 374-384.e6.

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Evaluating Hepatocyte Engraftment Efficiency

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The efficiency of Hep engraftment was evaluated every month via quantifying human ALB within mouse serum, using human Albumin ELISA Quantitation set (Catalog E80-129, Bethyl Laboratories, Montgomery, TX, USA) as per the manufacturer's instructions. Absorbance was read at 450 nm on a SpectraMax M3 (Molecular Devices).

Dagur R.S., Wang W., Cheng Y., Makarov E., Ganesan M., Suemizu H., Gebhart C.L., Gorantla S., Osna N, & Poluektova L.Y. (2018). Human hepatocyte depletion in the presence of HIV-1 infection in dual reconstituted humanized mice. Biology Open, 7(2), bio029785.

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Quantitative Liver Organoid Analysis

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Albumin production in 10 μL samples of LO media was measured using an enzyme linked immunosorbent assay (Human Albumin ELISA Quantitation Set, Bethyl laboratories Inc, Montgomery, TX) as per manufacturer’s instructions. Urea secretion was measured using the urease based colorimetric method described in Zawada et al. [17 (link)]. Glucose concentration in 10 μL samples was determined by the Yellow Springs Glucose 2300 STAT as per manufacturer’s instructions. Complete liver organoid growth medium (i.e. upcyte® Hepatocyte Growth Medium and upcyte® LSEC Growth Medium in 1:1 ratio) including all the adjuncts (e.g. FBS), was used as a blank for all readings.

Mattei G., Magliaro C., Giusti S., Ramachandran S.D., Heinz S., Braspenning J, & Ahluwalia A. (2017). On the adhesion-cohesion balance and oxygen consumption characteristics of liver organoids. PLoS ONE, 12(3), e0173206.

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Measurement of Autoantibody Levels in CSF and Sera

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Anti-Sm and anti-RNP levels in CSF and sera were measured using enzyme-linked immunosorbent assay (ELISA) kits, MESACUP™-3 test Sm and MESACUP™-2 test RNP (MBL, Nagano, Japan). Arbitrary unit was designated according to the manufacturer’s instructions. Anti-NR2 in sera and CSF was determined by specific ELISA using the highly purified synthetic 10 amino-acid peptide DWEYSVWLSN [6 (link)], conjugated to human serum albumin (HSA) as previously described [8 (link)]. Albumin levels in CSF and sera were measured by ELISA using the Human Albumin ELISA Quantitation Set (Bethyl Laboratories, Montgomery, TX, USA).

Hirohata S., Sakuma Y., Yanagida T, & Yoshio T. (2014). Association of cerebrospinal fluid anti-Sm antibodies with acute confusional state in systemic lupus erythematosus. Arthritis Research & Therapy, 16(5), 450.

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Quantitative Albumin Synthesis Assay

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The synthesis of albumin by hepatocytes was assessed using the Human Albumin ELISA Quantitation Set (Bethyl Laboratories) according to the manufacturer's instructions as previously reported.11

Figueiredo C., Oldhafer F., Wittauer E., Carvalho‐Oliveira M., Akhdar A., Beetz O., Chen‐Wacker C., Yuzefovych Y., Falk C.S., Blasczyk R, & Vondran F.W. (2019). Silencing of HLA class I on primary human hepatocytes as a novel strategy for reduction in alloreactivity. Journal of Cellular and Molecular Medicine, 23(8), 5705-5714.

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Quantifying Human Albumin in Microtissue

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The human albumin concentration of the harvested supernatants from single microtissues (24 hours after medium replenishment) was determined using the human Albumin ELISA Quantitation Set (E80-129/E101; Bethyl Laboratories, Inc., Montgomery, TX). The assay was performed according to the manufacturer's instructions.

Messner S., Fredriksson L., Lauschke V.M., Roessger K., Escher C., Bober M., Kelm J.M., Ingelman-Sundberg M, & Moritz W. (2018). Transcriptomic, Proteomic, and Functional Long-Term Characterization of Multicellular Three-Dimensional Human Liver Microtissues. Applied in Vitro Toxicology, 4(1), 1-12.

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Evaluating iPSC-Derived Hepatocyte Functions

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The ability of the iPSC-HLCs to both uptake LDL and secrete albumin was studied using the LDL Uptake Assay kit (Cayman Chemicals, USA) and Human Albumin ELISA Quantitation set (Bethyl Laboratories, USA), respectively, according to the manufacturers' instructions. Secreted albumin was measured from 24-h conditioned medium and normalised to the cell number.

Kiamehr M., Viiri L.E., Vihervaara T., Koistinen K.M., Hilvo M., Ekroos K., Käkelä R, & Aalto-Setälä K. (2017). Lipidomic profiling of patient-specific iPSC-derived hepatocyte-like cells. Disease Models & Mechanisms, 10(9), 1141-1153.

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Humanized-liver Chimeric Mice Characterization

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Humanized-liver chimeric TK-NOG mice were prepared as previously described^{27 (link)}. The human hepatocyte chimeric rate is correlated with serum human albumin levels, and the estimated human hepatocyte chimeric rate can be calculated from serum human albumin levels^{27 (link)}. Serum human albumin levels were measured with a Human Albumin ELISA Quantitation Set (Bethyl Laboratories Inc., Montgomery, TX, USA). All mouse experiments were conducted with the Guide for the Care and Use of Laboratory Animals from Osaka University Graduate School of Medicine and Central Institute for Experimental Animals (CIEA).

Murai K., Hikita H., Kai Y., Kondo Y., Fukuoka M., Fukutomi K., Doi A., Yamai T., Nakabori T., Fukuda R., Takahashi T., Miyakawa K., Suemizu H., Ryo A., Yamada R., Kodama T., Sakamori R., Tatsumi T, & Takehara T. (2020). Hepatitis C virus infection suppresses hepatitis B virus replication via the RIG-I-like helicase pathway. Scientific Reports, 10, 941.

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Albumin Quantification in CSF and Sera

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Albumin in CSF and sera was measured by ELISA using Human Albumin ELISA Quantitation Set (Bethyl Laboratories, Montgomery, Tx, USA).

Hirohata S., Arinuma Y., Yanagida T, & Yoshio T. (2014). Blood-brain barrier damages and intrathecal synthesis of anti-N-methyl-D-aspartate receptor NR2 antibodies in diffuse psychiatric/neuropsychological syndromes in systemic lupus erythematosus. Arthritis Research & Therapy, 16(2), R77.

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Quantifying Human Albumin Levels

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Human albumin levels were evaluated post-engraftment by using the human Albumin ELISA quantitation set (Bethyl Labs) as described by the manufacturer. For all assays, after incubation with the HRP-conjugated antibody, 3,3',5,5'-tetramethylbenzidine (TMB) peroxidase substrate (KPL) was applied, and the reactions were stopped by adding 2 N H₂SO₄ (Fisher Scientific) after 15–30 min. Plates were read at 450 nm on a VersaMax microplate reader (Molecular Devices).

Borel F., Tang Q., Gernoux G., Greer C., Wang Z., Barzel A., Kay M.A., Shultz L.D., Greiner D.L., Flotte T.R., Brehm M.A, & Mueller C. (2017). Survival Advantage of Both Human Hepatocyte Xenografts and Genome-Edited Hepatocytes for Treatment of α-1 Antitrypsin Deficiency. Molecular Therapy, 25(11), 2477-2489.

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