Amino Acid Analysis determines each amino acid's quantity in a protein, peptide, or free amino acid sample. The Bio-Synthesis's Amino Acid Analysis service provides quantitative information for 18 out of 20 standard amino acids except for tryptophan and cysteine. Tryptophan or cysteines are usually recovered in low quantities or destroyed during the analysis process. Asparagine is determined as aspartic acid and glutamine as glutamic acid.
Hydrolysis of proteins and peptides occurs using gas-phase HCl vapors in the presence of phenol. The released amino acid residues are analyzed on a UPLC AccQ·Tag Waters amino acid analyzer using a reversed-phase column to separate the released amino acids. Free amino acid samples are analyzed directly without the hydrolysis step.
Reports of results contain data chromatograms and tables of recovered amino acids.
Usually, recovered amounts are reported in micrograms, milligrams, or grams, depending on the sample amount analyzed. As an option, customers can provide by email (biosyn@biosyn.com) specific protein/peptide sequence to be compared with the experimental data.
Sample submission
Samples for amino acid analysis can be provided in liquid and in solid form.
Most samples containing proteins, peptides, and amino acids can be analyzed using amino acid analysis.
However, buffers with high amounts of primary and secondary amines, for example TRIS, need to be avoided, as well as high amounts of glycerin.
Avoid human and animal dandruff and dust in the sample.
Sample Amount
For samples in solution we recommend sending us >100 microliters of liquid sample at a minimum concentration of 0.1 microgram per microliter or 0.1 mg/ml. For best results provide samples dissolve in ultrapure water or PBS.
For solid sample, please provide enough to allow accurate weighing of samples. Usually, several mgs are required.
Please note:
Acid hydrolysis converts asparagine and glutamine to aspartic and glutamic acid, respectively. That is, the amino acid analysis result for Asp is a total of Asp + Asn and the result for Glu is Glu + Gln.
Please inquire for the analysis of unusual amino acids, such as hydroxyproline, taurine, Norleucine and Hydroxylysine.
Tryptophan Analysis
Tryptophan is destroyed by acid hydrolysis and requires a separate analysis using base hydrolysis or similar.
Cysteine Analysis
Quantitative determination of cysteine determination also requires a separate analysis. It is analyzed as cysteic acid after oxidation with performic acid. Please inquire or details.
Free Amino Acid Analysis
Free amino acid analysis is used to analyze samples that do not require hydrolysis such as amino acid supplements.
Shipment
For shipment place the sample into a microcentrifuge tube that can be locked!
1. Use a Safe Lock reaction tube or better, a borosilicate glass vial with a screw cap.
2. For solid samples, lyophilize or speed vac the protein to a solid sample to ensure protein stability during shipment.
3. Alternatively, for liquid samples refrigerate to +4°C for cold shipment of the liquid sample.
1. Download and fill out our Amino Acid Analysis Request Form.
2. Indicate if your primary objective is quantitation – for highest precision please supply protein sequence, accession number or expected composition, or
3. Your primary objective is amino acid composition, or
4. Your objective is both quantitation and composition.
The sample is now ready for submission
1. Download, fill out and sign the Amino Acid Sample Submission Form
2. Place the tube(s) in a bubble bag to protect the tube during shipment.
3. Ship the samples using a courier company. Liquid sample should be shipped at +4oC and gel samples at ambient temperature.
4. Ship to:
Bio-Synthesis Inc.,
Proteomic Analytical Division,
800 Mario Court,
Lewisville, TEXAS, 75057
USA
800.227.0627 (toll free)
https://www.biosyn.com/
https://www.biosyn.com/contactus.aspx
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