Product Details
| abID | ab65347 |
|---|---|
| Cat # +Size | K639-100 |
| Size | 100 assays |
| Detection Method | Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm) |
| Species Reactivity | Mammalian |
| Applications | N/A |
| Features & Benefits | • Simple procedure; takes ~ 30 minutes • Fast and convenient • Kit contains the necessary reagents for accurate measurement of L-Amino Acid in biological samples |
| Kit Components | • L-Amino Acid Assay Buffer • L-Amino Assay Probe (in DMSO) • L-Amino Acid Enzyme Mix • L-Amino Acid Standard (4nmol/µl) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
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. BioVision’s L-Amino Acid Assay Kit provides a convenient means for directly detecting L-amino acid in biological samples. There is no requirement for sample pretreatmen or purification when using this kit. 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.
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What is the way to prepare cell lysate samples?
For cell samples: Start with ~2 million 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 and use the supernatant 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.
Why is Glycine excluded from being detected in this kit?
Glycine, the simplest amino acid, has no enantiomers because it has two hydrogen atoms attached to the central carbon atom. Only when all four attachments are different can enantiomers occur.
Do different amino acids in the standard have different sensitivities when compared to each pure L-amino acid standard curve?
No, all the L-amino acids show identical sensitivities with the probe.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. It is recommended to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Li et al., Mitofusin-2-mediated tethering of mitochondria and endoplasmic reticulum promotes cell cycle arrest of vascular smooth muscle cells in G0/G1 phase. Acta Biochim Biophys Sin, Jun 2015; 47: 441 - 450.
Matsui et al., Dennd3 Functions as a Guanine Nucleotide Exchange Factor for Small GTPase Rab12 in Mouse Embryonic Fibroblasts. J. Biol. Chem., May 2014; 289: 13986 - 13995.
For more citations of this product click here
