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Add 60 µL of deionized water to the samples (dilute by a factor of 4). The Bicinchoninic Acid (BCA) Protein Assay primarily relies on two reactions. <>
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+?ͨ]�/-�N��m?����������,�r��G�F�b^�. These keywords were added by machine and not by the authors. This is a preview of subscription content. Since the production of Cu+in this assay is a function of protein concentration and incubation time, the protein content of unknown samples may be determined spectrophotometrically by comparison with known protein standards. The two assays are of similar sensitivity, but since BCA is stable under alkali conditions, this assay has the advantage that it can be carried out as a one-step process compared to the two steps needed in the Lowry assay. ;�w� �:;�|�Ld�Gd��*mr���ȟ~�����ϊ슞�{���g�f�����\�>����g�y��G��?� ���-���n�,�[�U!B6"U�����'�^���J~g��P�h�D^�L�*�u&d^�t��Uӽ�Eٛ�G˟�Gzv���\(���G��cP�\&1��%�F��dȨB�e�Ff��C����kt^V��fGE^���N�����!ҁ�OY��q���M����HdW��x:�J�W:��{��j�f��0�v~�6��H�X-�Bώ�~$je^�f����\�;y: 0000010404 00000 n
Part of Springer Nature. 23225 Pierce BCA Protein Assay Kit, sufficient reagents for 500 test-tube or 5000 microplate assays . endstream
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In my experience with cell lysates of tendon cells, one can expect to obtain about 0.4 ng of protein/cell. Clear, flat-bottom 96-well plate (such as Falcon, #353072), A plate reader that can measure absorbance at 560 nm, Typical lab supplies such as 1.5 mL Eppendorf tubes, cDNA strip tubes, pipettors, serological pipettes, micropipettors, multi-channel pipettor, reagent reservoirs, and pipette tips, Pipette 55 µL of BSA standard into tube 1 in a set of cDNA strip tubes, Pipette 55 µL of deionized water into tubes 2-8. The Rapid Gold BCA Protein Assay uses the same copper reduction method as the traditional BCA Protein Assay with a unique copper chelator. Analyt. endstream
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Schoel, B., Welzel, M., and Kaufmann, S. H. E. (1995) Quantification of protein in dilute and complex samples-modification of the bicinchoninic acid assay. 23225) or 500mL (in Product No. J. Biochem. %PDF-1.7
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July 2012. The BCA Protein Assay combines the protein-induced biuret reaction with the highly sensitive and selective colorimetric detection of the resulting cuprous cation (Cu1+) by bicinchoninic acid (BCA). Farhat, Y. The Enhanced Protocol requires a 60°C incubation for 30 minutes. Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., and Klenk, D. C. (1985) Measurement of protein using bicinchoninic acid. See data sheet.) 23225) or 500mL (in Product No. 159, 138–142. A purple colored reaction product is formed by the chelation of two molecules of BCA … 38 0 obj
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Your input will help make this resource better for everyone. 0000002921 00000 n
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, sodium bicarbonate, bicinchoninic acid and sodium tartrate in 0.1 M sodium hydroxide (Product No. �4R
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The Bicinchoninic Acid (BCA) Protein Assay is a highly sensitive colorimetric assay that is compatible with detergent solubilized protein solutions. 3. Analyt. endobj
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This process is experimental and the keywords may be updated as the learning algorithm improves. Youssef Farhat, Written 7/16/12, Last updated 5/6/14, Download the PDF or Microsoft Word versions of this protocol. 0000008654 00000 n
11) If your samples were diluted, be sure to scale back to the original concentration of the samples.