B-Phycoerythrin Protein
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Cat.No. : | B-Phycoerythrin-316 |
Product Overview : | B-Phycoerythrin Protein, activated. |
Description : | B-Phycoerythrin, activated was treated with a 15-fold molar excess of 2-Iminothiolane. Ratio SH-group/pigment: >= 3 (~5; Ellman’s reagent titration); Fluorescence: Emmax 576 nm (exc.: 542 nm in 0.1 M phosphate pH 7.2); |
Form : | 1 vial contains 1 mg pigment in 0.2 mL buffer (0.1 M phosphate, 5 mM EDTA) |
Applications : | Suitable for fluorescence. Activated B-Phycoerythrin is a reagent used to create B-Phycoerythrin type phycobiliproteins that may be analyzed via flow cytometry and immunofluorescence detection techniques. |
Notes : | Example for a labeling protocol: 1 mL purified at ~1.5 mg/mL in PBS (pH 7.5). Antibody must be fairly pure – i.e. purify on protein A or G column before conjugating. Add 20 µl SMCC, 4-(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester and incubate for 1 hour. Conjugation with activated phycobiliprotein: Immediately combine your activated phycobiliprotein with SMCC-Antibody. NOTE: For activated PE start with a 1:1 molar ratio. For activated APC, start with a 2:1 molar ratio. Incubate the mixture for 1-2 hours at room temperature. Block further reaction by adding 10:1 of NEM Solution(N-Ethylmaleimide) per mL of reaction (optional step). Incubate the mixture for 20 minutes at room temperature. Use a desalting column to exchange the conjugate into the desired storage buffer. To optimize the yield and size distribution of the conjugate, alter the amount of activated-Phycobiliprotein to your protein. |
Storage : | Store activated B-Phycoerythrin in the dark at 0 until -20centigrade. |
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