GGA3-PBD Beads (Binds active Arf1 and Arf6 proteins)

GGA3-PBD Beads (Binds active Arf1 and Arf6 proteins)
$0.00

Material

The GGA3-PBD containing residues 1-316 of GGA3 has been overexpressed in a bacterial expression system. The recombinant protein has a molecular weight of approximately 60kDa and contains a GST-tag at its amino terminus. The purified protein has been conjugated to colored sepharose beads and is supplied as a lyophilized purple powder (Figure 1). One tube of GGA07 should be sufficient for approximately 20-25 assays (see Biologcial Activity Assay on the product datasheet).

 

The GGA3-PBD protein used for conjugation to the beads was analysed by scanning densitometry of a Coomassie Blue stained 4-20% SDS polyacrylamide gel and determined to be >80% pure (see Figure 2)

 

The protein is supplied in a glutathione agarose bound format and is shipped lyophilized. The beads are colored for ease of use. This product is used in the Arf1 pull-down activation assay Biochem Kit™ (Cat. # BK032-S) and the Arf6 pull-down activation assay Biochem Kit™ (Cat. # BK033-S). 

beads
Figure 1. The beads provided are colored (a Cytoskeleton innovation in 2004) to easily identify pellet material.

Purity

A 20 µg sample of GGA3-PBD protein from two separate lots (molecular weight approx. 60 kDa) was separated by electrophoresis in a 4-20% SDS polyacrylamide gel, and stained with Coomassie Blue.  Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat. # ADV02).  See Blue molecular weight markers are from Invitrogen.   

Figure 2. Arf1 Activation Assay Biochem Kit™ Pull-down Assay Results.

gga07-bead-protein

Figure 2. A 20 µg sample of GGA3-PBD protein from two different lots (molecular weight approx. 60 kDa) was separated by electrophoresis in a 4-20% SDS polyacrylamide gel, and stained with Coomassie Blue.  Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat. # ADV02). See Blue  molecular weight markers are from Invitrogen. 

Biological Activity

The GGA3-PBD protein specifically recognizes and binds the active “GTP-bound” forms of the Arf1 and Arf6 proteins (1).  It has a much lower affinity for the inactive “GDP-bound” forms of Arf1 and Arf6.  When covalently coupled to a colored sepharose matrix, the GGA3-PBD beads become a convenient tool for assaying the activity of Arf1 and Arf6 proteins.  For standard biological assays using the beads, see product datasheet.

 

References

Takatsu, H., Yoshino, K., Toda, K., and Nakayama, K. (2002) GGA proteins associate with Golgi membranes through interaction between their GGAH domains and ADPribosylation factors. Biochem. J. 365, 369-378.

Coming soon!   If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com

Coming soon!   If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com