Human albumin elisa kit

Manufactured by Abcam

Sourced in United Kingdom, United States

The Human Albumin ELISA Kit is a quantitative assay designed to measure the concentration of human albumin in biological samples. It utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the target analyte.

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Spelling variants of this product

ELISAs (10 citations) Albumin ELISA kit (9 citations) Human albumin (2 citations) Human Albumin SimpleStep ELISA® Kit (2 citations)

38 protocols using human albumin elisa kit

Hepatocyte Differentiation and Function Assay

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Mature hepatocytes can secrete ALB and urea, therefore the concentrations of ALB and urea in the culture medium were detected. hADSCs and hUCMSCs were inoculated in 24-well plates at a density of 1×10
⁴ cells/mL. On the 7
^th and 15
^th day after induction, the culture medium of each group were collected and the ALB content was measured using the human albumin ELISA kit (ab108788; Abcam, Cambridge, UK) according to the manufacturer’s instructions. The absorbance was measured at a wavelength of 450 nm, and the concentration of ALB in the culture medium was calculated according to the standard curve. On the 7
^th and 15
^th day after induction, NH
₄Cl was added at a final concentration of 5 mM and the culture medium of each group were collected after 24 h of culture. The urea content in the medium was measured using the human urea ELISA Kit (ML07367; Shanghai Enzyme-linked Biotechnology, Shanghai, China) according to the manufacturer’s instructions. The absorbance was measured at a wavelength of 450 nm, and the concentration of urea in the culture medium was calculated according to the standard curve.

Jin Y., Shi R., Qi T., Li Q., Chen C., Gao S., Gao F., Yang D., Sun G., Xu J., Fu Q., Xu J, & Zhang X. (2023). Adipose-derived stem cells show hepatic differentiation potential and therapeutic effect in rats with acute liver failure: Adipose-derived stem cells for acute liver failure. Acta Biochimica et Biophysica Sinica, 55(4), 601-612.

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Quantifying Secreted Proteins in hEB Differentiation

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Conditioned medium coming from fully differentiated hEBs was collected and stored at −80 °C after 48 hours of the last change of medium. We then quantified albumin secreted from the differentiated embryoid bodies into the culture media using a Human Albumin ELISA kit (Abcam ab108788) following the manufacturer’s instructions. Alpha-fetoprotein secretion quantification assay was performed using an Alpha Fetoprotein Human SimpleStep ELISA kit (Abcam ab193765) according the manufacturer’s instructions. The quantification of Fibrinogen secretion into the culture supernatant was done using a Fibrinogen Human SimpleStep ELISA Kit (abcam – ab171578) following the manufacturer’s instructions. All the samples were carried out in triplicate.

Pettinato G., Lehoux S., Ramanathan R., Salem M.M., He L.X., Muse O., Flaumenhaft R., Thompson M.T., Rouse E.A., Cummings R.D., Wen X, & Fisher R.A. (2019). Generation of fully functional hepatocyte-like organoids from human induced pluripotent stem cells mixed with Endothelial Cells. Scientific Reports, 9, 8920.

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Albumin Production in Spheroid Cultures

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The Human Albumin ELISA Kit (Abcam, Cambridge, UK) was used to measure the albumin production by the spheroids from 48 h conditioned medium as a function of time, i.e., the measurements were performed for the same cell culture well at each time point to see the change in ALB levels from one time point to the next. In the spheroid experiment with vasculature, ALB levels were measured from two independent experiments, each containing three replicate wells.
From the chips, the medium samples were collected 24 h after the last medium change by pooling all three chip sites, i.e., six medium channels, from one chip into one at time points d8, (d14), and d20.

Suominen S., Hyypijev T., Venäläinen M., Yrjänäinen A., Vuorenpää H., Lehti-Polojärvi M., Räsänen M., Seppänen A., Hyttinen J., Miettinen S., Aalto-Setälä K, & Viiri L.E. (2023). Improvements in Maturity and Stability of 3D iPSC-Derived Hepatocyte-like Cell Cultures. Cells, 12(19), 2368.

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Functional Biomarker Assays of Hepatocytes

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Albumin, urea, and CYP450 enzyme assays were performed as functional biomarkers of the hepatocytes. Human Albumin ELISA Kit (cat# ab108787, Abcam, USA), Urea Assay Kit (cat# KA1652, Abnova, USA), and P450-Glo CYP3A4 Assay Kit (cat# V9001, Promega, USA) were used for albumin, urea, and CYP3A4 quantification, respectively. In brief, cell culture media samples were collected at specified time points and stored at − 80 °C. While CYP3A4 assay was performed by following the manufacturer’s instructions with slight modifications. Media samples were thawed at 37 °C in a water bath before experiments. A microplate reader (SpectraMax i3 Multimode Microplate Reader, Molecular Devices, USA) was used for taking readings by following the manufacturer’s instructions.

Farooqi H.M., Kang B., Khalid M.A., Salih A.R., Hyun K., Park S.H., Huh D, & Choi K.H. (2021). Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system. Nano Convergence, 8, 3.

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Quantifying Albumin Secretion Rates

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Culture media was collected after every change (n=3), diluted 1:10 in DPBS, and quantified according to manufacturer specifications using the human Albumin ELISA kit (Abcam). Absorbance was read on a Cytation3 Automated Microscope Plate Reader (BioTek). The Human Albumin ELISA has cross-reactivity with bovine albumin, and the albumin content of DMEM + 10% FBS was subtracted from all quantities. Cumulative albumin secretion was determined by summing all quantities and normalizing to cell number which was determined via dsDNA quantification described above.

Lewis P.L., Green R.M, & Shah R.N. (2018). 3D-Printed Gelatin Scaffolds of Differing Pore Geometry Modulate Hepatocyte Function and Gene Expression. Acta biomaterialia, 69, 63-70.

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Salivary Albumin Measurement Protocol

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Salivary albumin was measured for Cohort 1 using a purchased Human Albumin ELISA Kit (Abcam, ab108788). Assay was performed according to manufacturer instructions included with the kit.

Isho B., Abe K.T., Zuo M., Jamal A.J., Rathod B., Wang J.H., Li Z., Chao G., Rojas O.L., Bang Y.M., Pu A., Christie-Holmes N., Gervais C., Ceccarelli D., Samavarchi-Tehrani P., Guvenc F., Budylowski P., Li A., Paterson A., Yue F.Y., Marin L.M., Caldwell L., Wrana J.L., Colwill K., Sicheri F., Mubareka S., Gray-Owen S.D., Drews S.J., Siqueira W.L., Barrios-Rodiles M., Ostrowski M., Rini J.M., Durocher Y., McGeer A.J., Gommerman J.L, & Gingras A.C. (2020). Persistence of serum and saliva antibody responses to SARS-CoV-2 spike antigens in COVID-19 patients. Science Immunology, 5(52), eabe5511.

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Quantifying Albumin and Urea Secretion

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For albumin and urea quantification, media were collected at the end of the IHC stage and daily during the HM stage. For all experiments, the same volume of media was collected (10 mL). The level of albumin secreted in the livers was determined using a Human Albumin ELISA kit (Abcam, Cambridge, UK), following the manufacturer’s instructions. The urea nitrogen direct kit (Stanbio, Boerne, TX, USA) was used, following the manufacturer’s instructions, to determine the amount of urea secreted. Both albumin and urea contents were represented as micrograms secreted per liver.

Acun A., Oganesyan R., Jaramillo M., Yarmush M.L, & Uygun B.E. (2022). Human-Origin iPSC-Based Recellularization of Decellularized Whole Rat Livers. Bioengineering, 9(5), 219.

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Quantifying Secreted Human Albumin and Urea

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Spheroid medium supernatants of day 1, 3, 6, 9, 12, 15 and 18 were collected, centrifuged at 1250 rpm for 10 minutes to remove cell debris and stored at −20 °C until sample analysis. The amount of human serum albumin (HSA) secreted into the culture medium was determined by Human Albumin ELISA Kit (Abcam, UK) according to the manufacturer’s protocol. For ELISA, the supernatant was diluted 1:100 in dilution buffer. Urea was measured using a colorimetric assay kit from BioVision (Germany) according to the manufacturer’s protocol.

Eilenberger C., Rothbauer M., Ehmoser E.K., Ertl P, & Küpcü S. (2019). Effect of Spheroidal Age on Sorafenib Diffusivity and Toxicity in a 3D HepG2 Spheroid Model. Scientific Reports, 9, 4863.

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Albumin Secretion in Hepatocyte Models

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Cultured media from the HepG2 cells and human liver organoids were collected before and at 48 h after chemical treatment. The cells were incubated with 10 μM amikacin, amiodarone, sertraline, and 25 μM acetaminophen. The albumin content secreted into the culture medium was assayed using a human albumin ELISA kit (Abcam) in accordance with the manufacturer’s instructions. Data were reported as % value from the control that the cells no PL-inducer was treated.

Lee J.Y., Han H.J., Lee S.J., Cho E.H., Lee H.B., Seok J.H., Lim H.S, & Son W.C. (2020). Use of 3D Human Liver Organoids to Predict Drug-Induced Phospholipidosis. International Journal of Molecular Sciences, 21(8), 2982.

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Measuring Metabolic Activity and Secretion in Hepatocytes

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For measuring metabolic activity, PrestoBlue (Thermo Fisher Scientific) was added to culture media of rat hepatocytes on day 3 of culture at the concentration suggested by the manufacturer and incubated for 2 hours at 37°C, 10% CO₂. Following the incubation, media samples were used for the measurement.
For albumin and urea assays, the media collected from both rat and human hepatocytes at day 3 of culture was used. Level of albumin secreted in rat hepatocytes was determined following a direct, competitive enzyme-linked immunosorbent assay (ELISA) protocol as described before[25 (link)]. Albumin secreted by human hepatocytes was determined using Human Albumin ELISA kit (Abcam) following manufacturer’s instructions. The amount of urea secreted was determined using urea nitrogen direct kit (Stanbio) following the manufacturer’s instructions. The results were corrected to compensate for well-well differences in cell number using the cell counts.

Acun A., Oganesyan R., Uygun K., Yeh H., Yarmush M.L, & Uygun B.E. (2021). Liver donor age affects hepatocyte function through age-dependent changes in decellularized liver matrix. Biomaterials, 270, 120689.

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