Mouse serum concentration of hAlb, which is correlated with the repopulation index (27 (link)) was measured based on latex agglutination immunonephelometry (LX Reagent ‘Eiken’ Alb II; Eiken Chemical, Tokyo, Japan) as described previously (28 (link)).
Lx reagent eiken alb 2
Manufactured by Eiken Chemical
Sourced in Japan
The LX Reagent Eiken Alb II is a laboratory reagent used for the quantitative determination of albumin in human serum or plasma. It is designed to measure the concentration of albumin in a sample using a colorimetric method.
Automatically generated - may contain errors
Lab products found in correlation
Biomajesty series jca bm6050, by JEOL (1 mentions)
Dri chem 3500, by Fujifilm (1 mentions)
Transaminase cii test kit, by Fujifilm (1 mentions)
Bm6050 autoanalyzer, by JEOL (1 mentions)
Nuclease free water, by Thermo Fisher Scientific (1 mentions)
Smitest ex r d nucleic acid extraction kit, by Medical & Biological Laboratories (1 mentions)
Hbeag, by Abbott (1 mentions)
Collagenase, by Merck Group (1 mentions)
7 protocols using lx reagent eiken alb 2
1
Measuring Mouse Serum Albumin Levels
2
Serum Albumin Quantification Protocol
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Blood obtained was diluted in saline and albumin concentration measured by the clinical chemistry analyzer (BioMajesty™ Series JCA-BM6050, JEOL Ltd., Tokyo, Japan) using the latex agglutination immunonephelometry test (LX Reagent “Eiken” Alb II; Eiken Chemical Co., Ltd., Tokyo, Japan). Levels of human serum albumin were measured shortly after inoculation and then at weekly intervals.
3
Measuring Liver Toxicity in Transgenic Mice
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Blood was isolated from the tail vein of transgenic mice and serum was obtained by centrifugation. To select transgenic mice with liver toxicity due to the uPA transgene, alanine aminotransferase (ALT) activity was measured using the transaminase CII-test kit (WAKO Pure Chemical Industries, Ltd.). uPA and ALT activities in the selected transgenic mice were determined by ELISA (Mouse uPA Activity Assay, Molecular Innovations) and DRI-Chem 3500 (Fujifilm, Tokyo, Japan), respectively.
H-alb concentration in chimeric mouse blood was measured by immunonephelometry in a JEOL BM6050 autoanalyzer (JEOL, Tokyo, Japan) using LX Reagent Eiken Alb II (Eiken Chemical, Tokyo, Japan). Creatinine (CRE) and blood urea nitrogen (BUN) levels were measured by the urease glutamate dehydrogenase method and enzyme method using EKDIA XL ‘Eiken’ CREIII and EKDIA XL ‘Eiken’ UN (Eiken Chemical, Tokyo, Japan), respectively, with a JEOL BM6050 autoanalyzer (JEOL, Tokyo, Japan).
H-alb concentration in chimeric mouse blood was measured by immunonephelometry in a JEOL BM6050 autoanalyzer (JEOL, Tokyo, Japan) using LX Reagent Eiken Alb II (Eiken Chemical, Tokyo, Japan). Creatinine (CRE) and blood urea nitrogen (BUN) levels were measured by the urease glutamate dehydrogenase method and enzyme method using EKDIA XL ‘Eiken’ CREIII and EKDIA XL ‘Eiken’ UN (Eiken Chemical, Tokyo, Japan), respectively, with a JEOL BM6050 autoanalyzer (JEOL, Tokyo, Japan).
4
Generating Humanized Liver Chimeric Mice
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Chimeric mice with humanized liver (PXB-mice ® ) were generated by the transplantation of commercially available human hepatocytes (1-year-old, boy, Caucasian, BioIVT, Westbury, NY, USA) into cDNA-uPA/SCID mice (cDNA-uPA wild/+/SCID) (Yamasaki et al., 2020) . Hepatocytes (PXB-cells ® ) were isolated from male chimeric mice (15-22-week old) with blood human albumin (Alb) levels > 10 mg/mL (estimated replacement indexes > 90%) using a two-step collagenase perfusion method, as previously reported (Yamasaki et al., 2020) . Blood human albumin concentrations in PXB-mice ® were measured by immunonephelometry in a JEOL BM6050 autoanalyzer (JEOL, Tokyo, Japan) using LX Reagent Eiken Alb II (Eiken Chemical, Tokyo, Japan). The animal study protocol was approved by the Animal Care and Use Committee of PhoenixBio Co., Ltd. All experimental procedures were conducted in accordance with the guidelines provided by the Proper Conduct of Animal Experiments (June 1, 2006; Science Council of Japan).
5
Quantifying Human Hepatocyte Engraftment
6
Treating HBV in SCID-Hu Mice
7
Isolation of Humanized Hepatocytes
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All animal protocols described in this study were performed in accordance with the Guide for the Care and Use of Laboratory Animals 19 and approved by the Animal Welfare Committee of Phoenix Bio Co, Ltd (Hiroshima, Japan).
Cryopreserved HHs from four donors (Table 1) were purchased from BD Bioscience (San Jose, CA). By using these HHs, chimeric mice with humanized livers were prepared as described previously. 14 The human serum albumin (hAlb) concentration in mouse blood was measured at 3 and 6 weeks after transplantation and once per week thereafter, using latex agglutination immunonephelometry (LX Reagent Eiken Alb II; Eiken Chemical Co, Ltd, Tokyo, Japan) to estimate the replacement index of HHs in mouse livers. 20 The hAlb concentration in mouse blood correlated well with the replacement index determined on liver sections by immunohistochemistry using human-specific antibodies.
To isolate HHs, a total of 29 chimeric mice (donor 1, n Z 2; donor 2, n Z 11; donor 3, n Z 14; donor 4, n Z 2) with blood hAlb >12 mg/mL (replacement index of approximately 85%) were used. HHs were isolated using a two-step collagenase perfusion method using collagenase (Sigma Aldrich Japan, Tokyo), as previously described. 16
Cryopreserved HHs from four donors (Table 1) were purchased from BD Bioscience (San Jose, CA). By using these HHs, chimeric mice with humanized livers were prepared as described previously. 14 The human serum albumin (hAlb) concentration in mouse blood was measured at 3 and 6 weeks after transplantation and once per week thereafter, using latex agglutination immunonephelometry (LX Reagent Eiken Alb II; Eiken Chemical Co, Ltd, Tokyo, Japan) to estimate the replacement index of HHs in mouse livers. 20 The hAlb concentration in mouse blood correlated well with the replacement index determined on liver sections by immunohistochemistry using human-specific antibodies.
To isolate HHs, a total of 29 chimeric mice (donor 1, n Z 2; donor 2, n Z 11; donor 3, n Z 14; donor 4, n Z 2) with blood hAlb >12 mg/mL (replacement index of approximately 85%) were used. HHs were isolated using a two-step collagenase perfusion method using collagenase (Sigma Aldrich Japan, Tokyo), as previously described. 16
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