B-Phycoerythrin Protein

• Cat.No.: BPE-316
• Product Overview: B-Phycoerythrin Protein, activated.

Specification

• Tag: Non
• 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.