The extraction of the free amino acids was done as described by Hacham et al. (2002 (link)). Samples of 50 mg of frozen leaf powder were homogenized with 600 µl of water:chloroform:methanol (3:5:12 v/v/v). After centrifugation at 12,000 rpm for 2 min, the supernatant was collected and the residue was re-extracted with 600 µl of the same mixture, pooling the two supernatants. A mixture of 300 µl of chloroform and 450 µl of water were added to the supernatants, and after centrifugation the upper water:methanol phase was collected and dried in the SpeedVac. The samples were dissolved on 100 µl of sodium citrate loading buffer pH 2.2 (Biochrom, Holliston, MA, USA) and 10 µl were injected on a Biochrom 30 Amino Acid Analyser at the Protein Chemistry Service at CIB (CSIC, Madrid, Spain).
Sodium citrate loading buffer ph 2
Manufactured by Harvard Bioscience
Sourced in United States
Sodium citrate loading buffer pH 2.2 is a solution used to prepare samples for analysis in various laboratory techniques. It is a buffer solution with a pH of 2.2, primarily composed of sodium citrate. This buffer helps to maintain the sample's pH and ionic environment during preparation and analysis.
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Lab products found in correlation
3 protocols using sodium citrate loading buffer ph 2
1
Free Amino Acid Extraction from Leaves
2
Quantitative Analysis of Leaf Amino Acids
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The extraction of free amino acids was done as described by Hacham et al. (2002) (link). Samples of 50 mg of frozen leaf powder were homogenized with 600 μl of water:chloroform:methanol (3:5:12 v/v/v). After centrifugation at 12,000 rpm for 2 min, the supernatant was collected and the residue was re-extracted with 600 μl of the same mixture, pooling the two supernatants. A mixture of 300 μl of chloroform and 450 μl of water were added to the supernatants, and after centrifugation the upper water:methanol phase was collected and dried in the SpeedVac. The samples were dissolved on 100 μl of sodium citrate loading buffer pH 2.2 (Biochrom, United States) and 10 μl were injected on a Biochrom 30 Amino Acid Analyser (Biochrom, United States) at the Protein Chemistry Service at CIB (CSIC, Madrid, Spain).
3
Amino Acid Extraction and Analysis
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The extraction of the free amino acids was done as described by [28 (link)]. Samples of 50 mg of leaf frozen powder were homogenized with 600 μl of water:chloroform:methanol (3:5:12 v/v/v). After centrifugation at 12000 rpm for 2 min, the supernatant was collected and the residue was re-extracted with 600 μl of the same mixture, pooling the two supernatants. A mixture of 300 μl of chloroform and 450 μl of water were added to the supernatants, and after centrifugation the upper water:methanol phase was collected and dried in a SpeedVac. The samples were dissolved on 100 μl of sodium citrate loading buffer pH 2.2 (Biochrom, USA) and 10 μl were injected on a Biochrom 30 Amino Acid Analyser (Biochrom, USA) at the Protein Chemistry Service at CIB (CSIC, Madrid, Spain).
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