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Control Peptide, PB1 |
 | | | | Control Peptide, PB1 | | | Discontinued as of 4/1/09. Contact info@neb.com for assistance. | |
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This product is not available for online ordering.
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Description:
Control Peptide, PB1 is a synthetic peptide containing an N-terminal cysteine (CTRSRHSSYPNEYEEDEEMEEEL); the sequence is derived from mouse Bad. This peptide can be ligated to the C-terminus of the thioester tagged Carrier Protein 39 (NEB #'s E6602 or E6603) or Carrier Protein 27 (NEB #'s E6606 or E6607) (1-3) based on a procedure known as intein-mediated protein ligation (IPL), resulting in a native peptide bond between the reactants (4-6). PB1 is included as a control peptide in the Peptide-Carrier™ Kit (NEB #E6600) and is also sold separately.
Figure 1: Western blot analysis of antiserum raised against Control Peptide, PB1. PB1 was synthesized with a N-terminal cysteine (CTRSRHSSYPNEYEEDEEMEEEL; MW 2862 Da) and ligated to Carrier Protein 39 (Western) (NEB #E6602). The reaction times and temperatures are indicated. The samples were electrophoresed on a 12% SDS polyacrylamide gel and transferred to nitrocellulose. Anti-PB1 Peptide Antibody (NEB #E6608) was used as the primary antibody. Lanes 1 and 5 are controls (C) which contain only Carrier Protein 39 (Western).
Figure 2: Analysis of peptide ligation by Coomassie Blue stained SDS-PAGE. The Control Peptide, PB1, was ligated to Carrier Protein 39 (Western), as in ProtocoI II on the Control Peptide, PB1 (NEB #S6605) datacard. The ligation samples were electrophoresed on a 12% SDS polyacrylamide gel and examined by Coomassie Blue staining. Lanes 1 and 5 are controls (C) which contain only Carrier Protein 39 (Western).
Source:
A HPLC purified synthetic peptide dissolved in water to a concentration of 1 mM.
Properties & Usage
Molecular Weight:
Theoretical: 2,862 daltons
Reaction & Storage Conditions
Concentration:
1 mM
Storage Temperature:
-70°C
Avoid repeated freeze/thaw cycles.
Notes
General notes:- The sample can be stored at -20°C after the addition of 3X SDS Sample Buffer. For enzymatic assays Protocol II on the Control Peptide, PB1 (NEB #S6605) datacard can be used to produce the substrates: adjust substrate concentration as recommended. In this case do not add 3X SDS Sample Buffer.
- Can be stored for 1 month or less at -20°C. Should be stored at -70°C upon arrival.
Usage notes:- 75-90% of the Carrier Protein 39 (Western) is ligated when the purified peptide is used at a final concentration of 0.5-1 mM.
Use of certain unpurified peptides may result in lower ligation efficiency (10-50%). This will still result in a positive signal on Western blot analysis. - We recommend the use of Control Peptide, PB1 at 0.5 mM final concentration.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Analysis:
Carrier Protein 39 (Western) is ligated to the Control Peptide, PB1 containing an N-terminal cysteine (CTRSRHSSYPNEYEEDEEMEEEL; MW, 2862 Da) for 60 minutes at room temperature. A ligated product is detected by Coomassie Blue staining of a SDS-PAGE.
References
- Sun, L., Rush, J., Ghosh, I., Maunus, J.R. and Xu, M.-Q. (2004) Biotechniques, 37.
- Ghosh, I., Sun, L., Evans. T.C., Jr. and Xu, M.-Q. (2004) J. Immunol. Methods, In press.
- Xu, J., Sun, L., Ghosh, I. and Xu, M.-Q. (2004) Biotechniques, 36, 976-981.
- Evans, T.C., Jr., Benner, J. and Xu, M.-Q. (1998) Protein Sci., 7, 2256-2264.
- Muir, T.W., Sondhi, D. and Cole, P.A. (1998) Proc. Natl. Acad. Sci. USA, 95, 6705-6710.
- Evans, T.C., Jr. and Xu, M.-Q. (1999) Biopolymers, 51, 333-342.
Companion Products
Anti-PB1 Peptide Antibody
Carrier Protein 27 (Array)
Carrier Protein 27 (Western)
Carrier Protein 39 (Array)
Carrier Protein 39 (Western)
Peptide-Carrier™ Kit
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