What can interfere with BCA?
What can interfere with BCA?
The presence of reducing agents such as dithiothreitol (DTT), ß-mercaptoethanol and TCEP at concentrations routinely used in protein sample buffers can interfere with the modified Lowry and BCA protein assays.
Does EDTA interfere with BCA?
EDTA and reducing agents, such as DTT, BME, TCEP and other disulfide reducing agents, interfere with the BCA assay. The 2-mercaptoethanol will interfere with the BCA assay, and the SDS will interfere with the Bradford assay.
Is BCA compatible with SDS?
Note: As used in this protocol, 5% SDS is compatible with the BCA Protein Assay.
What are the limitations of the Bradford assay?
The main limitation of the Bradford assay is its incompatibility with most detergents, routinely used to solubilize membrane proteins. (Interestingly, however, very low levels of non-ionic detergent, such as Triton X-100, may improve sensitivity and variability of the Bradford assay [25] ).
Is BCA compatible with detergent?
The advantages offered by the assay help to explain the popularity and widespread use over other protein assays. The advantages of the BCA include: Compatible with detergent concentrations up to 5%
Is BCA compatible with urea?
5% SDS and 3M urea are compatible with BCA assay. It is compatible with detergents and urea.
Is BCA better than Bradford?
Historically, the BCA method is more sensitive than the Bradford method, because the first method is based on protein-copper chelation and secondary detection of the reduced copper. Whereas the Bradford method is based on protein-dye binding and colour shift from 465 to 595 nm.
What is the difference between BCA and Bradford?
The key difference between BCA and Bradford assay is that BCA assay is time-consuming and less accurate, whereas Bradford assay is quick and accurate. BCA and Bradford assay are two assay methods of protein concentration determination.
Does peg interfere with BCA assay?
PEG doesn’t chelate copper nor is a reducing agent so your BCA results should be fine.
Is BCA more sensitive than Bradford?
Why is BSA used in BCA assay?
Typically, standard solutions of bovine serum albumin (BSA) are used to produce a calibration curve of absorbance versus mass concentration. Assuming the analyte-proteins react in the same manner as the BSA standard, the unknown concentration can be determined.
What is BCA chemical?
The Bicinchoninic Acid Assay (BCA), also referred to as the Smiths’ Assay, is a biochemical assay designed by Paul Smith in 1985. (Smith, et al., [7]) This assay uses colorimetric detection and quantification to determine the total concentration of protein in a solution by exhibiting a measurable color change.
What metals should not be used with Zincalume® steel or colibond® steel?
Due to a phenomenon known as galvanic corrosion, some commonly used metals can cause accelerated corrosion when used with ZINCALUME® steel or COLORBOND® steel. Copper and lead are two such metals. Copper pipes and lead flashing should not come in direct contact with either ZINCALUME® steel or COLORBOND® steel.
How to mix BCA and copper together?
Combine 50 parts BCA solution with 1 part Copper solution, for example, for 10ml working solution combine 10ml BCA solution with 0.2ml Copper Solution. The mixed Working Solution should be a clear, green solution.
What is the absorbance of copper and BCA at 550 nm?
The BCA/copper complex is water-soluble and exhibits a strong linear absorbance at 562 nm with increasing protein concentrations. The purple colour can be measured at any wavelength between 550 nm and 570 nm with minimal (less than 10%) loss of signal.
What are the advantages and disadvantages of BCA reagent?
The advantage of BCA is that the reagent is fairly stable under alkaline conditions, and can be included in the copper solution to allow a one-step procedure. Substances that reduce copper will also produce colour in the BCA assay, thus interfering with the accuracy of the protein quantitation.