Albumin Secretion by hepatocytes into culture medium was measured every 24 h using Rat Albumin ELISA Quantitation Kit from Bethyl Laboratories, Inc [Montgomery, TX] according to manufacturer’s instructions. In short, a 96 well plate was coated with a coating antibody for 1 hour and blocked with BSA for 30 min. Standard/Sample was added to each well and incubated for 1 hour. HRP detection antibody was incubated for 1 hour, followed by the addition of TMB Substrate solution that was developed in the dark for 15 min. Absorbance was read on AD340 plate spectrophotometer [Beckman Coulter, Brea, CA] at 450 nm. Data were expressed as the means ± SD from six independent experiments.
Rat albumin elisa quantitation kit
Manufactured by Fortis Life Sciences
Sourced in United States
The Rat Albumin ELISA Quantitation Kit is a laboratory equipment product designed to quantify albumin levels in rat samples. It utilizes the enzyme-linked immunosorbent assay (ELISA) technique to measure the concentration of albumin present in the sample.
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Lab products found in correlation
Ad340 plate spectrophotometer, by Beckman Coulter (2 mentions)
P ace mdq, by Beckman Coulter (1 mentions)
Rat tnf α platinum elisa kit, by Thermo Fisher Scientific (1 mentions)
Quant ittm picogreen dsdna assay kit, by Thermo Fisher Scientific (1 mentions)
Urea nitrogen kit, by EKF Diagnostics (1 mentions)
Spectramax plus reader, by Molecular Devices (1 mentions)
Creatinine companion kit, by Exocell (1 mentions)
Albuwell m elisa kit, by Exocell (1 mentions)
10 protocols using rat albumin elisa quantitation kit
1
Albumin Secretion by Hepatocytes
2
Induction and Measurement of Mesangioproliferative Glomerulonephritis in Rats
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Mesangioproliferative glomerulonephritis (GN; anti-Thy1.1 nephritis) was induced by a single intravenous injection of a 1.75-mg/kg OX-7 monoclonal antibody (mAb) into the tail vein [49 (link)] or respective sham treatment. Nephritic rats were kept in metabolic cages to determine urinary albumin excretion in 24 h. The rat albumin ELISA quantitation kit (Bethyl Laboratories, Montgomery, USA) was set up using Nunc-Immuno 96-well flat-bottom high-binding MaxiSorp polystyrene microtiter plates (Nunc, Roskilde, Denmark) to quantitatively measure levels of urinary albumin [49 (link)]. Only rats exhibiting marked increases of urinary albumin were regarded as animals with successful GN induction [49 (link)]. After 1 week, renal neurons were harvested and kidneys prepared for further investigations.
3
Hepatocyte Albumin Secretion Dynamics
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Albumin secretion from cultured hepatocytes was measured over the four days of cultivation. On HB-EGF affinity-bound heparin-IC (HB-EGF 76.2 ng/cm2) and collagen type I coated TCPS (collagen-TCPS; Iwaki Glass, Chiba, Japan), hepatocytes were cultured in a culture medium with or without soluble EGF (10 ng/mL). Hepatocytes on PIPAAm were cultured in culture medium containing both soluble heparin (1.0 μg/cm2) and soluble HB-EGF (100 ng/cm2) with or without soluble EGF (10 ng/mL). Albumin secreted into the culture medium was collected at 24 h intervals and quantified with the rat albumin ELISA Quantitation kit (Bethyl Laboratories, Montgomery, TX, USA).
4
Quantifying Hepatocyte Function Markers
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Alb secretion was determined by using the Rat Albumin ELISA Quantitation Kit (Bethyl Laboratories Inc., Montgomery, USA). Urea synthesis by hepatocytes incubated in fresh medium containing 2 mM NH4Cl for 90 min was measured with the Urea Nitrogen kit (BioChain Institute Inc., Boerne, USA). CYP1A enzymatic activity and UGT were assessed by the conversion of 50 mM CEC and 100 mM 4-MU (Sigma-Aldrich, USA) to CHC and 4-methylumbelliferyl glucuronide (4-MUG) in 60-min incubations, respectively. CHC and 4-MUG were detected at an excitation wavelength of 325 nm as previously described using capillary electrophoresis (P/ACE MDQ, Beckman Instruments Inc., USA).
5
Quantifying Inflammatory Markers and Cell Viability
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All chemicals were, unless otherwise stated, of analytical grade and obtained from Sigma-Aldrich (Madison, WI, USA). The Rat TNFα Platinum ELISA kit was obtained from eBioscience (San Diego, CA, USA), Rat Albumin ELISA Quantitation kit was from Bethyl Laboratories (Montgomery, TX, USA). Kit for measurement of lactate dehydrogenase activity was obtained from DiaSys (Holzheim, Germany), substrate for WST-1 test was obtained from Roche (Penzberg, Germany). Fluorescence probe dichlorodihydrofluorescein diacetate (DCFDA) was obtained from Life Technologies (Carlsbad, CA, USA).
6
Quantifying Urinary Protein Excretion
7
Urinary Albumin Quantification in Rats
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Rats were kept in metabolic cages 24 h, 48 h, and 6 days after antibody administration for determination of urinary albumin and urinary protein excretion during 24 h. The rat albumin ELISA Quantitation Kit (Bethyl Laboratories, Montgomery, USA) was set up using Nunc-Immuno 96-well flat-bottom high-binding Maxisorb-polystyrene microtiter plates (Nunc, Roskilde, Denmark) to quantitatively measure levels of urinary albumin [58 (link)].
8
Quantifying Hepatocyte Function Metrics
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Albumin production was measured using a rat albumin ELISA quantitation kit (Bethyl laboratories Inc., USA). Urea synthesis was measured using a urea nitrogen kit (Stanbio laboratory, USA). For CYP1A2 activity analysis, CYP specific probe substrate (200 μM phenacetin) was diluted in Krebs-Henseleit buffer (KHB) and incubated with the cells for 1.5 hours. The supernatant were collected and stored at −80 °C until being measured by liquid chromatography-mass spectrometry (LC/MS). All functional data were normalized to one million cells by using a Quant-iTTM picoGreen dsDNA assay kit (Invitrogen, Singapore).
9
Quantification of Albumin Secretion
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Albumin secretion by hepatocytes into culture medium was measured every 24 h using the rat albumin ELISA quantitation kit from Bethyl Laboratories, Inc. (Montgomery, TX, USA) according to the manufacturer’s instructions. In short, a 96 well plate was coated with a coating antibody for 1 h and blocked with BSA for 30 min. The standard/sample was added to each well and incubated for 1 h. HRP detection antibody was incubated for 1 h, followed by the addition of TMB substrate solution that was developed in the dark for 15 min. Absorbance was read on AD340 plate spectrophotometer (Beckman Coulter, Brea, CA, USA) at 450 nm. Data were expressed as the means ± SD from six independent experiments.
10
Quantifying Urine Albumin in Rodents
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For rats, albumin was analyzed using the Rat Albumin Elisa Quantitation Kit (Bethyl Laboratories Inc., Montgomery, TX) and corrected for the urine creatinine concentration measured with the Jaffé reaction using a creatinine standard solution (Sigma, St Louis, MO). For mice, albumin was analyzed using the Albuwell M ELISA Kit and all urine samples were corrected for the urine creatinine concentration measured with the Creatinine Companion Kit (both from Exocell Inc., Philadelphia, PA). Optical density was measured on a Spectra max plus reader (Molecular Devices, Sunnyvale, CA).
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