Overview

  • Product name

    L-Amino Acid Assay Kit
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay type

    Quantitative
  • Sensitivity

    <= 40 µM
  • Assay time

    0h 30m
  • Product overview

    L-Amino Acid Assay Kit ab65347 is used measure the level of L-amino acids in biological samples. There is no requirement for sample pretreatment or purification.


    The L-amino acid(s) level can be quantified using fluorometric (at Ex/Em = 535/587 nm) or colorimetric (at λ = 570 nm) methods in 96-well plates.


    Amino acid assay protocol summary:
    - add reaction mix to sample and standard wells
    - incubate for 30 min
    - analyze with microplate reader

  • Notes

    L-Amino acids are the most essential elements in biology. Accurately quantitating L-amino acids in body fluids or purified samples may provide valuable information for diagnostic or basic research studies.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    L-Amino Acid Assay Buffer WM 1 x 25ml
    L-Amino Acid Enzyme Mix Green 1 vial
    L-amino Acid Probe Red 1 x 0.2ml
    L-Amino Acid Standard (4 nmol/µl) Yellow 1 x 300µl
  • Research areas

  • Relevance

    Amino acids perform various important functions for the body and serve as the materials for the body's cells, hormones, and enzymes. Of the standard amino acids, all by glycine can exist in either of the two optical isomers, called D and L enantiomers, which are mirror images of each other. The L-amino acids represent the vast majority of amino acids found in proteins during translation in the ribosome, and are therefore the most essential element in biology and metabolic functions.

Images

  • Standard curve (fluorimetrical): mean of duplicates (+/-) with background subtracted

  • Standard curve (colorimetrical): mean of duplicates (+/-SD) with background readings subtracted

  • L-Amino Acid Standard Curve.

Protocols

References

This product has been referenced in:

  • Modvig IM  et al. Peptone-mediated glucagon-like peptide-1 secretion depends on intestinal absorption and activation of basolaterally located Calcium-Sensing Receptors. Physiol Rep 7:e14056 (2019). Read more (PubMed: 31020803) »
  • Posada IMD  et al. Opposite feedback from mTORC1 to H-ras and K-ras4B downstream of SREBP1. Sci Rep 7:8944 (2017). Read more (PubMed: 28827765) »
See all 4 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer


ab83374 (Branched Chain Amino Acid Assay Kit). We have not tested this kit with cell or tissue lysates yet but only with fluids like Cell culture supernatant, Urine, Serum, Plasma and can therefore not provide more than a general protocol below.

ab65347 (L-Amino Acid Assay Kit) has not been tested with cell lysates but the 96 well format is for liquid samples. Please use 50 ul samples per well. There is not need to grow the cells in the small wells.

The following general guidelines can be followed:

For cell samples: Start with ˜2X106 cells, suspend the cell pellet 500 μl (or ˜4 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

For tissue samples: Start with 10-100 mg of the tissue, add 500-1000 μl (or ˜4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

Read More

Answer

Merci de nous avoir contactés.

Le laboratoire m'a confirmé que nous n'avons malheureusement pas de données expérimentales pour ces kits sur des lysats musculaires.

Désolé de ne pas pouvoir vous aider davantage. N'hésitez pas néanmoins à nous contacter de nouveau si vous avez d'autres questions.

Read More

Answer


I am afraid that this is proprietary information which cannot be disclosed.

We apologize for any inconvenience this may cause.

Read More

Answer

I general, werecommend using several doses of your serum sample to ensure the readings are within the standard curve range. In essence, the dose range selection depends on your expectation of the L-amino-acid concentration in your samples.

An example of a range of serum volumes you may try is : 0, 1, 5, 10, 25 and 50ul/well.

I hope this is helpful. Please contact us again if you have any further questions.

Read More

Question
Answer

Thank you for your enquiry.

I am sorry to confirm we do not currently have specific data in terms of what reagents and at what concentrations would interfere with this assay. Theoretically chemicals at high concentrations and ionic detergents have to be avoided when the assay is done. Ifyou are concerned of using specific chemicals in the assay, then you could consider optimizing the concentrations at which they are compatible with the assay.

We aim to provide as much information as possible to customers, and I am sorry we have no more specific details to provide to help answer your question on this occasion. If you have any further questions, please do not hesitate to contact us.

Read More

Question
Answer

Thank you for your phone call today. I have gotten in touch with the production laboratory, and I was told that the kit only detects free L-amino acids. I hope this is helpful but please let me know if you have further questions.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up