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Succinic acid assay kit

Manufactured by Megazyme
Sourced in Ireland, United States

The Succinic Acid Assay Kit is a laboratory equipment used to quantify the concentration of succinic acid in a sample. It provides a rapid and accurate method to measure succinic acid levels.

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3 protocols using succinic acid assay kit

1

Measuring Serum Succinate Levels in Rats

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Serum levels of succinate were monitored in a separate cohort of rats raised from weaning with or without amoxicillin (50 mg/kg body weight) until euthanization at time point 2. Animals were euthanized by CO2 inhalation. Blood was collected through heart puncture and serum was separated and stored at −80°C. Stored serum was thawed and diluted 1:4 times with distilled water and quantitated for succinate using the Succinic Acid Assay Kit from Megazyme (catalog no.: K‐Succ; Megazyme Ltd, Bray, Ireland; Figure 11).
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2

Succinic Acid Production in Methanotrophic Bacteria

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Example 3

Positive transformants of M. capsulatus containing the malate dehydrogenase (mdh; SEQ ID 1; pSB107), pyruvate carboxylase (pyc; SEQ ID 2, pSB108), phospho enol pyruvate carboxylase (ppc; SEQ ID 3, pSB109) genes were verified by PCR. These were inoculated into 5 ml of liquid NMS media taken in 30 ml culture tubes and sealed with suba seals. 15 ml of Methane mixture (95% CH4; 5% CO2) was introduced into the culture tube using a syringe. The tubes were incubated at 45° C. at 250 rpm agitation. Once the culture OD reached 1, the cultures was centrifuged and the supernatant samples were taken and assayed for succinic acid. The organic acid concentrations were measured using Succinic Acid assay kit (Megazyme International; K-SUCC) according to manufacturer's protocol (FIG. 3).

Some of the recombinant strains had higher levels of succinic acid compared to the control strain. Recombinant strain with SEQ ID 1 was >15% improved when compared to the native strain. Recombinant strain with SEQ ID 2 was >50% improved when compared to the native strain.

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3

Metabolite Dynamics in Cell Cultures

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The levels of the metabolites acetate, glucose, lactate, and succinate were measured in the time-course samples (0, 2, 6, 10, 24, 48, 72 h) of vcDMEM and ccDMEM, as well as the endpoint sample VF (72 h) using the Acetate Colorimetric Assay Kit (MAK086, Sigma, USA), the Glucose-Glo TM Assay kit (J6021, Promega, USA), the Lactate-Glo TM Assay kit (J5021, Promega, USA), and the Succinic Acid Assay kit (K-SUCC, Megazyme, Germany) according to the manufacturers’ protocols. Measurements were made on an EnSpire 2300 plate reader (PerkinElmer Life Sciences, Schwerzenbach, Switzerland). Acetate, glucose, lactate, and succinate concentrations were calculated by linear regression based on the standard (0.00 to 10.00 nmol for acetate, 4.39 to 50.00 mM for glucose, 0.20 to 50.00 mM for lactate, and 0.13 µg to 4.00 µg total succinate) after subtraction of the respective enzyme blanks. Biological triplicates were analyzed for each sample and median and range (min, max) were calculated to represent the measured values. Linear reduction/accumulation rates were calculated using linear regression analysis in R and expressed as concentration changes in mM per hour.
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