TRAF-6 Inhibitor Peptide Set

Functions as a TRAF6 decoy by binding to the T6DP motif of RANK, thereby preventing binding of RANK to TRAF6.

The TRAF6 inhibitory peptide also contains a protein transduction (PTD) sequence (DRQIKIWFQNRRMKWKK) derived from antennapedia which renders the peptide cell permeable. The control peptide consists of only the PTD sequence.

Preparation of 5 mM Stock Solutions
PBS* is added directly to the vials to prepare the stock solutions. Note: Bring the solution to room temperature and quick spin the tubes before opening the caps.

TRAF6 Inhibitor Peptide: 1 mg of DRQIKIWFQNRRMKWKKRKIPTEDEY
Add 57.7 ul of PBS* to the vial to make a 5 mM stock solution. Mix by vortexing. Aliquot and store at -20C or -80C. Avoid repeated freeze thawing.

Control Peptide: 1 mg of DRQIKIWFQNRRMKWKK
Add 84.8 ul PBS* to the vial. Mix by vortexing. Aliquot and store at 20C or -80C. Avoid repeated freeze thawing.


*Recipe for 1X PBS:
1. Dissolve the following in 800ml distilled H2O.
- 8g of NaCl
- 0.2g of KCl
- 1.44g of Na2HPO4
- 0.24g of KH2PO4
2. Adjust pH to 7.5 with HCl.
3. Adjust volume to 1L with additional distilled H2O.
4. Sterilize by autoclaving

TRAF6 Inhibitor peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKKRKIPTEDEY (TRAF6 binding sequence: RKIPTEDEY). Molecular weight: 3494. Antennapedia Control peptide: 2 x 1 mg (lyophilized) DRQIKIWFQNRRMKWKK. Molecular weight: 2361.

TRAF6

Inhibition of RANKL mediated osteoclast differentiation.

Researchers can study the effect of TRAF6 inhibitor peptide using a variety of methods, such as EMSA, NF-kB/p65 ELISA,IkBa phosphorylation ELISA, osteoclast differentiation assay. The Osteoclast formation assay protocol is a guideline which may need to be optimized for different cell types.