Alanine aminotransferase activity assay kit

Manufactured by Merck Group

Sourced in United States

The Alanine Aminotransferase Activity Assay Kit is a colorimetric assay that measures the activity of the enzyme alanine aminotransferase (ALT). ALT is an enzyme found in various tissues, particularly the liver. The kit provides the necessary reagents to quantify ALT activity in biological samples such as serum, plasma, or cell lysates.

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Lab products found in correlation

Aspartate aminotransferase activity assay kit, by Merck Group (3 mentions) Balb c scid mice, by Jackson ImmunoResearch (2 mentions) Matrigel, by Corning (2 mentions) Colorimetric aspartate aminotransferase activity assay kit, by Merck Group (2 mentions) Cobase 501, by Roche (1 mentions) Enspire multimode plate reader, by PerkinElmer (1 mentions) Colorimetric assay kit, by Merck Group (1 mentions) Aspartate aminotransferase ast activity assay kit, by Merck Group (1 mentions) Ezrmi 13k, by Merck Group (1 mentions) Bilirubin assay kit, by Merck Group (1 mentions) Microtainer blood collection tube, by BD (1 mentions) Flexstation 3, by Molecular Devices (1 mentions) Elisa kit, by Abcam (1 mentions) High sensitivity glucose assay kit, by Merck Group (1 mentions)

13 protocols using alanine aminotransferase activity assay kit

Therapeutic Antibody Evaluation in Breast Cancer Xenografts

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Therapy experiments were carried out using protocols approved by the
Texas A&M University IACUC. BALB/c SCID mice were purchased from Jackson
Laboratories, bred and maintained in specific pathogen-free housing. 6-8-week
old female mice were implanted with 3-5 × 10⁶ MDA-MB-453 cells
or 4-5 × 10⁶ JIMT-1 cells suspended in 100 μl 50%
RPMI1640 media and 50% Matrigel (Corning, Corning, NY, USA) in the mammary fat
pad. When the tumor size reached a volume of 50-100 mm³, mice were
randomized into groups and dosed intravenously with 2 mg/kg antibody, ADC, or
vehicle twice with a three week interval (MDA-MB-453) or four times with one
week intervals (JIMT-1). Tumor size was determined using the formula:

tumor size = \frac{tumor length \times {tumor width}^{2}}{2}

, where the tumor length and tumor width were
measured with digital calipers every 3-4 days for the duration of the
experiment. When tumor volumes reached a size of ~2000 mm³,
mice were euthanized. Alanine aminotransferase activity in serum samples from
mice were analyzed using Alanine Aminotransferase Activity Assay Kits
(Sigma-Aldrich) and the manufacturer’s protocol.

Kang J.C., Sun W., Khare P., Karimi M., Wang X., Shen Y., Ober R.J, & Ward E.S. (2019). Engineering a HER2-specific antibody-drug conjugate to increase lysosomal delivery and therapeutic efficacy. Nature biotechnology, 37(5), 523-526.

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Therapeutic Antibody Evaluation in Breast Cancer Xenografts

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tumor size = \frac{tumor length \times {tumor width}^{2}}{2}

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Hepatocellular Injury Evaluation with ALT

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Hepatocellular injury was assessed using plasma samples in duplicate at 1:2 dilution with an Alanine Aminotransferase Activity Assay kit according to the manufacturer’s protocol (Sigma-Aldrich).

Keeler G.D., Gaddie C.D., Sagadevan A.S., Senior K.G., Côté I., Rechdan M., Min D., Mahan D., Poma B, & Hoffman B.E. (2022). Induction of antigen-specific tolerance by hepatic AAV immunotherapy regardless of T cell epitope usage or mouse strain background. Molecular Therapy. Methods & Clinical Development, 28, 177-189.

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Quantifying Blood Lipid and Liver Enzymes

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Total cholesterol (Chol), high-density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C) contents were assessed. As a measurement of liver damage, serum alanine (ALT) activity was also quantified. Enzymatic colorimetric kits (Roche Diagnostics GmbH; Mannheim, Germany) were used to measure these parameters in a Roche HITACHI Cobas e501 instrument (Roche Diagnostics GmbH; Mannheim, Germany) accordingly to manufacturer’s instructions by enzymatic colorimetric assay (Roche Diagnostics GmbH; Mannheim, Germany). ALT/AST activity in cell supernatants upon the treatments described before, were assessed by Alanine Aminotransferase Activity Assay Kit and Aspartate Aminotransferase Activity Assay Kit (Sigma-Aldrich^®, St. Louis, MO, USA). The amount of pyruvate and glutamate quantified with these colorimetric kits are respectively proportional to ALT and AST enzymatic activity in each sample. The assay and the kinetic measurements were performed following manufacturer’s instructions. The absorbance for ALT was measured at 570 nm and for AST at 450 nm using an EnSpire^® Multimode Plate Reader (Perkin Elmer).

Marin V., Gazzin S., Gambaro S.E., Dal Ben M., Calligaris S., Anese M., Raseni A., Avellini C., Giraudi P.J., Tiribelli C, & Rosso N. (2017). Effects of Oral Administration of Silymarin in a Juvenile Murine Model of Non-alcoholic Steatohepatitis. Nutrients, 9(9), 1006.

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Evaluating Liver Injury Biomarkers

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After euthanasia of animals by carbon dioxide (CO2) inhalation using a rodent euthanasia unit (Open Science, AE0904), and then blood was collected from animals in vacutainers with EDTA from the unpaired posterior vena cava (vena cava posterior). The extent of liver injury was determined via measuring the serum level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), which are commonly used markers of liver injury [19 (link)]. The levels of the two markers were determined using commercially available kits AsAT-Vital kit (Vital Diagnostics SPb, Russia), and Alanine aminotransferase Activity Assay Kit (sigma-Aldrich), respectively. Another measured marker of hepatitis injury was total cholesterol (TC) and triglycerides (TG), which was determined using the colorimetric Assay Kit (Sigma-Aldrich, St. Louis, MO, USA). The levels of cytokines were analyzed in blood serum, according to manufacturer's instructions, from specific rat ELISA kits for TNF-α, IL-6 and IFN- γ (Sigma-Aldrich, St. Louis, MO, USA).

Sergazy S., Shulgau Z., Kamyshanskiy Y., Zhumadilov Z., Krivyh E., Gulyayev A, & Aljofan M. (2023). Blueberry and cranberry extracts mitigate CCL4-induced liver damage, suppressing liver fibrosis, inflammation and oxidative stress. Heliyon, 9(4), e15370.

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Hepatic Triglyceride and Liver Enzyme Assays

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Concentrations of hepatic triglycerides were measured in hepatic homogenates with a colorimetric assay (no. 10010303, Cayman Chemical) according to the protocol recommended by the manufacturer
Concentrations of ALT (MAK052-Sigma Aldrich, Alanine Aminotransferase Activity Assay Kit) and AST (MAK055-Sigma Aldrich, Aspartate Aminotransferase Activity Assay Kit) in serum were determined with a colorimetric coupled enzymatic assay according to the procedure recommended by the manufacturer

Hill C.M., Albarado D.C., Coco L.G., Spann R.A., Khan M.S., Qualls-Creekmore E., Burk D.H., Burke S.J., Collier J.J., Yu S., McDougal D.H., Berthoud H.R., Münzberg H., Bartke A, & Morrison C.D. (2022). FGF21 is required for protein restriction to extend lifespan and improve metabolic health in male mice. Nature Communications, 13, 1897.

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Serum Analysis of B16F10 Tumor-Bearing Mice

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Serum was isolated from B16F10-bearing mice 3d after 1x AIP treatment. AST and ALT levels were quantified by using a colorimetric aspartate aminotransferase activity assay kit (Sigma Aldrich) or alanine aminotransferase activity assay kit (Sigma Aldrich), respectively, according to the manufacturer’s protocol.

Wang C., Cui A., Bukenya M., Aung A., Pradhan D., Whittaker C.A., Agarwal Y., Thomas A., Liang S., Amlashi P., Suh H., Spranger S., Hacohen N, & Irvine D.J. (2021). Reprogramming NK cells and macrophages via combined antibody and cytokine therapy primes tumors for elimination by checkpoint blockade. Cell reports, 37(8), 110021.

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Quantifying Liver Enzyme Levels

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Serum was isolated from mice 36 hours after treatment and liver enzymes AST and ALT were quantified using a colorimetric Aspartate Aminotransferase Activity Assay Kit (Sigma Aldrich) and Alanine Aminotransferase Activity Assay Kit (Sigma Aldrich) respectively according to the manufacturer’s protocol.

Moynihan K.D., Opel C.F., Szeto G.L., Tzeng A., Zhu E.F., Engreitz J.M., Williams R.T., Rakhra K., Zhang M.H., Rothschilds A.M., Kumari S., Kelly R.L., Kwan B.H., Abraham W., Hu K., Mehta N.K., Kauke M.J., Suh H., Cochran J.R., Lauffenburger D.A., Wittrup K.D, & Irvine D.J. (2016). Eradication of large established tumors in mice by combination immunotherapy that engages innate and adaptive immune responses. Nature medicine, 22(12), 1402-1410.

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Serum Analysis of Insulin and Liver Enzymes

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At 25 weeks, blood was obtained by cardiac puncture under deep naesthesia using isoflurane. Serum was separated by centrifugation (3000 rpm, 10 min). Enzyme-linked immunosorbent assays for mouse insulin (EZRMI-13K; Millipore, Boston, MA, USA) were undertaken according to the manufacturer’s instructions. ALT activity was measured using an alanine aminotransferase activity assay kit (Sigma, St. Louis, MO, USA). Serum aspartate aminotransferase activity was measured using an aspartate aminotransferase (AST) activity assay kit (Sigma, St. Louis, MO, USA) according to the manufacturer’s instructions.

Ren J., Wang X., Yee C., Gorrell M.D., McLennan S.V, & Twigg S.M. (2022). Sitagliptin Is More Effective Than Gliclazide in Preventing Pro-Fibrotic and Pro-Inflammatory Changes in a Rodent Model of Diet-Induced Non-Alcoholic Fatty Liver Disease. Molecules, 27(3), 727.

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Serum Biomarker Measurement Protocol

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Blood samples were collected using BD Microtainer® blood collection tubes (BD, Cat#365967) and stored at −80°C. Serum levels of total bilirubin were determined with a Bilirubin Assay Kit (Sigma-Aldrich, MAK126), and alanine aminotransferase levels with an Alanine Aminotransferase Activity Assay Kit (Sigma-Aldrich, MAK052).

Airik M., McCourt B., Ozturk T.T., Huynh A.B., Zhang X., Tometich J.T., Topaloglu R., Ozen H., Orhan D., Nejak-Bowen K., Monga S.P., Hand T.W., Ozaltin F, & Airik R. (2022). Mitigation of portal fibrosis and cholestatic liver disease in ANKS6-deficient livers by macrophage depletion. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 36(2), e22157.

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