BioVision’s Extra Sense™ BCA Protein Assay kit provides an ultra-sensitive method to detect and quantify total protein concentration even in the presence of detergents. The Kit has been optimized for determining total protein content in low protein concentration samples (0.5-40 μg/ml). The assay is based on the reaction of bicinchoninic acid (BCA) with the cuprous cation (Cu+1), which is generated by reduction of cupric cation (Cu+2) with the protein in alkaline conditions. The Cu+1-BCA chelate is a water-soluble complex and exhibits a strong absorbance at 562 nm that is linear over a concentration range of 0.5-40 μg/ml. In general, protein concentrations are estimated with reference to a commonly used protein standard. The Kit includes Bovine Serum Albumin (BSA) as a protein standard for estimation of total protein content of samples.
|Cat # +Size||K814-2500 (2500 assays)
K814-5000 (5000 assays)
|Detection Method||Absorbance (562 nm)|
|Features & Benefits||• optimized for determining total protein content in low protein concentration samples (0.5-40 μg/ml)|
|Kit Components||• BCA Reagent A • BCA Reagent B • BCA Reagent C • BSA (2 mg/ml)|
|USAGE||For Research Use Only! Not For Use in Humans.|
Can I use cell culture supernatant as a sample with this ELISA kit?
Yes, you can use cell culture supernatant as a sample.
Could you please explain the principle clearly? Also, does the size of my fusion protein matters in the detection process?
1. E4700 is a competitive ELISA kit, where the FLAG peptide is coated to the 96 well plates and the antibody (against Flag protein) that is provided with the kit will bind to the FLAG-tagged protein in the sample and will be removed during the wash steps. The remaining antibody (unbound) will bind to the wells. This means that the higher the concentration of your FLAG protein, less unbound antibody will bind to the wells, leading to lower OD readings and vice versa. Basically, the competition is between the bound and unbound antibody.
2. Regarding your question whether the size of the fusion protein matters, it is not the size, but it is the amount of the fusion protein that matters. The primary antibody is going to be the limiting factor and the primary antibody included in the kit is enough and not going to be saturated.
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