We measured urine Klotho using Klotho ELISA kits (Immuno-Biological Laboratories Co, Tokyo, Japan). We measured urine NGAL using NGAL ELISA kits (R&D Systems, Inc., Minneapolis, MN, USA). Samples were used with one freeze/thaw cycle and were detected by a technician with a blind method. The intra- and inter-assay coefficient of variation for Klotho were both< 15% and for NGAL were both< 10%. The results were corrected for urine creatinine excretion.
Ngal elisa kit
Manufactured by R&D Systems
Sourced in United States
The NGAL ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed to measure the levels of Neutrophil Gelatinase-Associated Lipocalin (NGAL) in various sample types. NGAL is a protein involved in immune response and is often used as a biomarker for certain medical conditions. This kit provides a standardized and reliable method to detect and quantify NGAL concentrations.
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Lab products found in correlation
Liblia hfabp latex agglutination turbidimetric immunoassay, by Sumitomo Pharma (1 mentions)
Zag enzyme linked immunosorbent assay elisa kit, by BioVendor (1 mentions)
Cystatin c elisa kit, by R&D Systems (1 mentions)
Apoptosis detection kit, by Merck Group (1 mentions)
Il 1β elisa kit, by Abcam (1 mentions)
Mcp 1 elisa kit, by Abcam (1 mentions)
Tnf α elisa kit, by R&D Systems (1 mentions)
Bun assay kit, by Nanjing Jiancheng (1 mentions)
7 protocols using ngal elisa kit
1
Urine Klotho and NGAL Measurement
2
Biomarker Analysis for Kidney Injury
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Main equipment consisted of a microplate reader (Jiangsu Potebio Biotechnology Co., Ltd., Jiangsu, China), automatic biochemical instrument (Precise, Beijing, China), centrifuge (Beijing Guangan Medical Equipment Factory), pipettes (Dragon Laboratory Instruments Ltd., Beijing, China), EP tubes and centrifuge tubes (Haimen Innovative Experimental Equipment Factory, Jiangsu, China). Experimental reagents consisted of NGAL ELISA kits (R&D Systems, Inc., Minneapolis, MN, USA), CysC kits (Beijing Strong Biotechnologies, Inc., Beijing, China) and SCr kits (BioSino, Beijing, China).
3
Biomarker Measurement in Acute Kidney Injury
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The urine and blood samples were collected within 30 min of admission. The urine and blood samples were centrifuged within 5 min at 4°C and immediately frozen at −80°C until they were analysed. The serum levels of HFABP, hsTnT, NT‐proBNP, and BNP were measured on admission. In addition, the urine NGAL, LFABP, β2MG, NAG, and albumin excretion were also measured on admission. These urine and serum biomarkers were measured by the Special Reference Laboratory (SRL©, Tokyo, Japan). The level of urine LFABP was measured with an enzyme‐linked immunosorbent assay (ELISA) using a human LFABP ELISA Kit (Kyowa Medex Co., Tokyo, Japan). The level of urinary NGAL was measured using the NGAL ELISA Kit (R&D Systems, Inc., Minneapolis, MN). The serum HFABP was measured using a MARKIT‐M HFABP ELISA Kit until June 2012 and using a LIBLIA HFABP latex agglutination turbidimetric immunoassay from July 2012 (DS Pharma Biomedical, Osaka, Japan). The lower and upper limits of detection for the urinary NGAL concentration were 4 and 500 pg/mL, respectively, and the lower limit for the urine LFABP was 2.9 pg/mL.
4
Biomarkers in Cirrhosis Progression
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In the groups without ascites and with ascites but without treatment, serum samples were collected when patients presented at the hospital. In the group with ascites and treatment, serum collection was performed immediately before the administration of tolvaptan. Albumin (g/dL), total bilirubin (mg/dL), sodium (mEq/L), creatinine (mg/dL), BUN (mg/dL), CRP (mg/dL), α-fetoprotein (AFP; ng/mL) and des-γ-carboxy prothrombin (DCP; mAU/mL) were measured. Serum samples were kept at -80 °C until copeptin (pmol/L), ZAG (μg/dL), cystatin C (mg/dL), NGAL (ng/mL) and L-FABP (ng/mL) measurements were performed using an automated copeptin immunofluorescent assay kit (Thermo Fisher Scientific Inc., Tokyo, Japan), ZAG enzyme-linked immunosorbent assay (ELISA) kit (BioVendor, Brno, Czech Republic), cystatin C ELISA kit (R&D Systems, Minneapolis, MN, USA), NGAL ELISA kit (R&D Systems) and high-sensitivity human L-FABP ELISA kit (CMIC Holdings, Tokyo, Japan), respectively.
The Child-Pugh score, albumin-bilirubin (ALBI) score (18 (link)), fibrosis index based on 4 factors (FIB-4) (19 (link)), model for end-stage liver disease (MELD) score, MELD-Na score and estimated glomerular filtration rate (eGFR) with Cockcroft and Modification of Diet in Renal Disease (MDRD) formula were calculated.
The Child-Pugh score, albumin-bilirubin (ALBI) score (18 (link)), fibrosis index based on 4 factors (FIB-4) (19 (link)), model for end-stage liver disease (MELD) score, MELD-Na score and estimated glomerular filtration rate (eGFR) with Cockcroft and Modification of Diet in Renal Disease (MDRD) formula were calculated.
5
Kidney Injury Biomarkers in Blood and Urine
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Creatinine and urea nitrogen levels were detected in blood and urine samples. Paraffin sections were used for in situ end labelling (ISEL) of fragmented DNAs with digoxigenindeoxyuridine by terminal deoxynucleotidyl transferase using an Apoptosis Detection Kit (Millipore, Billerica, MA, USA). Apoptotic cells were examined at 400 × magnification over 20 fields of tubulointerstitial areas and semi-quantitatively scored18 (link). The expression of neutrophil gelatinase-associated lipocalin (NGAL) was measured in both serum and urine using the NGAL ELISA kit (R&D Systems, Minneapolis, MN, USA).
6
Cytokine and Biomarker Profiling
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The cultured medium of NRK-52E cells and the serum samples of the experimental mice were collected and analyzed using the MCP-1 ELISA kit (Abcam, Cat. No. ab208979) and IL-1β ELISA kit (Abcam, Cat. No. ab197742). The serum samples were also analyzed using TNF-α ELISA kit (R&D Systems, Cat. No. MTA00B) and neutrophil gelatinase-associated lipocalin (NGAL) ELISA kit (R&D Systems, Cat. No. MLCN20) in accordance with the manufacturer’s instructions.
7
Biomarkers of Early Renal Injury
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Blood urea nitrogen (BUN) and neutrophil gelatinase-associated lipocalin (NGAL) were used as biomarkers to estimate early renal injury in the current study. The concentration of BUN was measured by a commercial BUN assay kit (Nanjing Jiancheng Bioengineer Institute, Nanjing, China), and NGAL was measured by an NGAL ELISA kit (R&D Systems, USA). Experimental procedures in the provided instruction manuals were strictly followed.
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