محلول برادفورد 5X حجم 50 سی سی DNAbiotech

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محلول برادفورد 5X حجم 50 سی سی DNAbiotech

Bradford Reagent, 5X

Cat #: DB9701-50ml

Volume: 50 ml

Sensitivity: (aprox.) 20 ugr - 1.5 mgr/ml

Package: Dark brown glass bottle

For Molecularr Biology research

Storage: 4^o C

www.kalazist.ir

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Bradford Reagent

Notes:

1. The change in the color of Coomassie G-250 from red to blue upon binding protein is measured spectroscopically. In the absence of protein, when the dye is red, Bradford reagent has an absorbance maximum (A_max) of 470 nm.
2. A standard curve, also known as a calibration curve, is a type of graph used as a quantitative research technique. Multiple samples with known properties are measured and graphed, which then allows the same properties to be determined for unknown samples by interpolation on the graph 
3. BSA is used because of its stability to increase signal in assays, its lack of effect in many biochemical reactions, and its low cost, since large quantities of it can be readily purified from bovine blood, a byproduct of the cattle industry.

   Many protein-containing solutions have the highest absorption at 280 nm in the spectrophotometer, the UV range. This requires spectrophotometers capable of measuring in the UV range, which many cannot. Additionally, the absorption maxima at 280 nm requires that proteins contain aromatic amino acids such as tyrosine (Y), phenylalanine (F) and/or tryptophan (W). Not all proteins contain these amino acids, a fact which will skew the concentration measurements. If nucleic acids are present in the sample, they would also absorb light at 280 nm, skewing the results further. By using the Bradford protein assay, one can avoid all of these complications by simply mixing the protein samples with the Coomassie Brilliant Blue G-250 dye (Bradford reagent) and measuring their absorbances at 595 nm, which is in the Vis range.

   The procedure for Bradford protein assay is very easy and simple to follow. It is done in one step where the Bradford reagent is added to a test tube along with the sample. After mixing well, the mixture almost immediately changes to a blue color. When the dye binds to the proteins through a process that takes about 2 minutes, a change in the absorption maximum of the dye from 465 nm to 595 nm in acidic solutions occurs. This dye creates strong noncovalent bonds with the proteins, via electrostatic interactions with the amino and carboxyl groups, as well as Van Der Waals interactions. Only the molecules that bind to the proteins in solution exhibit this change in absorption, which eliminates the concern that unbound molecules of the dye might contribute to the experimentally obtained absorption reading. This process is more beneficial since it is less pricey than other methods, easy to use, and has high sensitivity of the dye for protein

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