|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.|