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Description
Protein quantification is one of the important steps in protein research such as Western blot assay. Protein assay methods are diverse, and there is one which absorbs at 280nm without special reagents or operation. This method is simple, yet it also has a shortcoming not being able to analyze protein assay when no amino acid such as phenylalanine, tryptophan or tyrosine are present. Neither will it be used commonly due to its DNA absorbance interference (AI). The other methods are Lowry assay or BCA assay, which are highly sensitive for protein quantification, but is quite inconvenient and time-consuming. Contrarily, Bradford method is the most commonly used one (within 10min), which can detect the minimum amount of protein and is very simple to use. This PRO-MEASURE is even more convenient than Bradford assay, declining AI of solution itself so that the background absorbance is maintained very low.
Characteristics
1) Very Simple Usage steps
2) Less than 10 minutes for measurement
3) High sensitivity to detect 10-25ng/ml of protein
4) Lower Background interference
Kit Contents
* PRO-MEASURETM Solution (10X) 100㎖
* Standard Solution (BSA, 1mg/㎖) 1㎖
Storage : 4 C; (stable at least 1 year)
Remarks
1) In protein quantification, there can be an error due to high AI, depending on the solution type used for protein assay. Therefore, one mus t perform calibration using a blank (background absorbance) to nullify possible errors. For example, it has to be handed carefully if using a solution containing detergents such as Triton X-100, NP-40, deoxycholate etc., for they effect the absorbance of protein assay. iNtRON PRO-PREP (Cat. No.17081) can extract protein without affecting the protein assay.
2) In protein assay, one must use the original solution for dilution. To keep an error to minimum, one can perform dilution by using the protein extracting solution or using sterile water if necessary.
3) When measuring absorbance, use plastic cuvette. Because, this solution can coat the surface of glass cuvette. As the results, this solution give effect for absorbnce measurement. PRO-MEASURE solution on glass cuvette surface can be removed by MtOH washing.
4) When quantification of protein, normally assay in duplicate or triplicates. Because we can decrease an measurement error.
5) For each protein measurement, one should calculate standard curve. By the way, the equation of iNtRON is suitable for any experiment with relative equal volume (Western blot, etc).
6) When measuring absorbance at 595nm, you can use cuvette or 96wellmicro plate.
7) Suitable reaction time is within 2-10 min after the addition of samples. Absorbance will increase over time, solution should incubate at room temperature for no more than 1 hour.
Cat. # 21011
Created on 01/04/2005 02:10 AM by admin
Updated on 08/28/2005 07:48 AM by admin
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