B-Phycoerythrin (B-PE) is a light-harvesting pigment protein that can be isolated from the red alga Porphyridium cruentum. The absorption spectrum of B-Phycoerythrin peaks at 545 nm and 563 nm, with a shoulder at 498 nm [1] [2].
In Vitro, Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs). 1. Preparation of B-Phycoerythrin (B-PE) This product is stored in 60% ammonium sulfate solution containing EDTA and sodium azide and needs to be desalted before use. Centrifuge B-Phycoerythrin at 10,000 g for 10 minutes at 4°C, discard the supernatant, and resuspend the precipitate (B-Phycoerythrin) in the desired buffer (usually 1X PBS (pH 7.2-7.4). Desalt the sample using Sephadex G-25 or dialysis, repeat at least 2 times. 2. Activation of B-Phycoerythrin After desalting, you can modify and activate the algal protein with the appropriate bioconjugation reagent based on your own samples for conjugation with the target protein. 3. Antibody preparation (0-16 hours) Purified antibody solution at a concentration of 2-10 mg/mL (4 mg/mL or higher is recommended for best results) and does not contain BSA or other proteins. Most buffers are acceptable (e.g. MES, phosphate and Tris buffers, with or without NaCl, can all be used successfully) as long as the pH is between 6-8 and there are no oxidants or other compounds that may interfere with antibody reduction. The exact time depends on the protocol used. 4. Calculation of the degree of antibody labeling (DOL) For antibodies, the optimal degree of labeling (DOL) is usually between 2-10. The value depends mainly on the properties of the label and the antibody/protein, so it needs to be gradually determined through multiple small-scale labeling experiments.
References:
[1]. Glazer A N, et al. Subunit structure and chromophore composition of rhodophytan phycoerythrins. Porphyridium cruentum B-phycoerythrin and b-phycoerythrin[J]. Journal of Biological Chemistry, 1977, 252(1): 32-42.
[2]. Munier M, et al. Physicochemical factors affecting the stability of two pigments: R-phycoerythrin of Grateloupia turuturu and B-phycoerythrin of Porphyridium cruentum[J]. Food chemistry, 2014, 150: 400-407.
In Vitro, Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs). 1. Preparation of B-Phycoerythrin (B-PE) This product is stored in 60% ammonium sulfate solution containing EDTA and sodium azide and needs to be desalted before use. Centrifuge B-Phycoerythrin at 10,000 g for 10 minutes at 4°C, discard the supernatant, and resuspend the precipitate (B-Phycoerythrin) in the desired buffer (usually 1X PBS (pH 7.2-7.4). Desalt the sample using Sephadex G-25 or dialysis, repeat at least 2 times. 2. Activation of B-Phycoerythrin After desalting, you can modify and activate the algal protein with the appropriate bioconjugation reagent based on your own samples for conjugation with the target protein. 3. Antibody preparation (0-16 hours) Purified antibody solution at a concentration of 2-10 mg/mL (4 mg/mL or higher is recommended for best results) and does not contain BSA or other proteins. Most buffers are acceptable (e.g. MES, phosphate and Tris buffers, with or without NaCl, can all be used successfully) as long as the pH is between 6-8 and there are no oxidants or other compounds that may interfere with antibody reduction. The exact time depends on the protocol used. 4. Calculation of the degree of antibody labeling (DOL) For antibodies, the optimal degree of labeling (DOL) is usually between 2-10. The value depends mainly on the properties of the label and the antibody/protein, so it needs to be gradually determined through multiple small-scale labeling experiments.
References:
[1]. Glazer A N, et al. Subunit structure and chromophore composition of rhodophytan phycoerythrins. Porphyridium cruentum B-phycoerythrin and b-phycoerythrin[J]. Journal of Biological Chemistry, 1977, 252(1): 32-42.
[2]. Munier M, et al. Physicochemical factors affecting the stability of two pigments: R-phycoerythrin of Grateloupia turuturu and B-phycoerythrin of Porphyridium cruentum[J]. Food chemistry, 2014, 150: 400-407.