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Protocol: Prepare working solution: Mix reagent A (in general the blue bottle in a BCA kit) and reagent B with ratio of A:B=1:20 for enough volume of using. Wells with a ratio value of less than 1.7 are considered to be positive for the presence of protein. 2. Briefly, 25 μL of protein standard or sample are added to the micro-well followed by the addition of 200 μL BCA working reagent (1:8). A. Microplate Procedure: 1. 5.2 Microplate Assay Protocol 1. 4 Preparation of working reagent Add 20 µl of reagent S to each ml of reagent A that will be needed for the run. Pipette working solution into a microplate … The microplate is sealed and allowed to incubate at 37 °C for 30 minutes. Background absorbance of the microplate is removed by subtraction of a blank well. �LNƩn8�'�j�y/���z5�g�+���0\k�,�����ׁ��c���Pc1����c���uޯknZ=�q���EHf��?���?�. Do In the microplate protocol of the BCA assay, the sample volume is up to 25 µl. Both assays were conducted according to the respective manufacturers’ protocols, in a microplate format. The BCA assay was performed in a 96-well microplate as per the manufacturers’ recommendations. 23227 BCA Protein Assay Reagent Kit, sufficient reagents for 250 test tube or 2,500 microplate assays Kit Contents: 23228 BCA Reagent A, 500 ml, containing sodium carbonate, ... Dilution Scheme for Standard Test Tube Protocol and Microplate Procedure (Working Range = 20–2,000 µg/ml) ��EF��V�U(�������eu��������fv��������7GWgw��������8HXhx��������)9IYiy��������*:JZjz���������� �� ? Known concentrations were based on manufacturers’ indicated concentrations and confirmed by absorbance at 280 nm. �b�+��"m��c��c�i�S52�Dž����S*���J�H��/�q�q�v����g���ȸ��b������9�8�����ʥs����q�q�q�f�8p�F2���U�����*��LP.a\a��]1�s�Ԃ���ym*Ҩ#��V2;�&4�Ec�� v.�}�ǩ�3S*�T� �p\i������j���1�r��eq6]�(�Tv��.Jp4�9���R �
��8���{o�z����LB�WS-�; Add 25 µl of each BSA Standard and protein samples into microtiter plate wells. VII. ����� If precipitate forms, warm the solution and vortex. This is the protocol to be used in the determination of total protein in brain tissue by the Pierce BCA method. %PDF-1.2
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#"""#''''''''''�� � �" �� � Protocol name: 260_280 Ratio.prt The protocol calls for an absorbance measurement at 260 nm and 280 nm. Assay Protocol: BCA Assay can be performed in a microtiter plate format or test tube format. (This working reagent A' is stable for 1 week even though a precipitate will form after 1 day. Using a Transformation the A 260/A 280 ratio is calculated. Add 200 µl of BCA working reagent to the Standard & sample wells, mix thoroughly for 30 s. 3. Microplate Protocol: 20-2,000 g/ml The Micro BCA ... No microplate protocol is currently available Requires heating for color development Interfering Substances6 Compatible with all reducing agents and detergents found at concentrations routinely used in protein sample buffers. ���� Adobe d� �� C Figure 4. Accuracy of the Pierce Rapid Gold BCA Protein Assay and Bradford Protein Assay with known protein mixes.
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