Glucose Regulated Protein 94 (Grp94, gp96)

Grp94 (glucose regulated protein 94, gp96) is a constitutively expressed endoplasmic reticulum (ER) lumenal protein that is up-regulated in response to cellular stress such as heat shock, oxidative stress or glucose depletion. Grp94 is thought to play a role in protein translocation to the ER, in their subsequent folding and assembly, and in regulating protein secretion\n(1). Grp94 also plays a role in antigen presentation by accessing the endogenous pathway and eliciting specific CTL responses to chaperone bound peptides via MHC class I pathway (2) Grp94 is a member of the Hsp90 family of stress proteins and shares sequence homolgy with its cytosolic equivalent, Hsp90 (3). Both Hsp90 and Grp94 are calcium binding proteins (4). Despite sharing 50% sequence homology over its N domains and complete conservation in its ligand binding domains with Hsp90, Grp94 and Hsp90 differ in their interactions with regulatory ligands as Grp94 has weak ATP binding and hydrolyisis activity (5). Grp94 exists as a homodimer and the two subunits interact at two distinct intermolecular sites, C terminal dimerization domains and the N-terminal interacts with the middle domain of opposing subunits. (6). Grp94 contains a carboxy terminal KDEL (Lys-Asp-Glu-Leu) sequence which is believed to aid in its retention in the ER (7).\n\nApplications:\nSuitable for use in Western Blot, Immunoprecipitation and Flow Cytometry. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 0.5ug/ml was sufficient for detection of Grp94 in 20ug of HeLa lysate; Goat anti-rat IgG:HRP as the secondary antibody.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

G3057-40B

Size

50ug

Applications

FC, IP, WB

Hosts

Rat

Reactivities

Hum, Mouse, Rat, Bov, Can, Ch/Bird, GP, NHP, Prc, Rabbit, Sheep, Xen/Amph

Form

Supplied as a liquid in PBS, pH 7.2, 0.09% sodium azide and 50% glycerol.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG2a

References

1. Rudolph, R.W., and Bedows, E., (1997) J Biol Chem 272:3125-3128. 2. Srivastava, P.K., et al., (1994) Immunogenetics 39(2):93-98. 3. Mazzarella, R.A., and Green M. (1987) J Biol Chem 262:8875-8883. 4. Kang, H.S. and Welch, W.J., (1991) J Biol Chem 266(9): 5643-5649. 5. Soldano, K.L., et al., (2003) J Biol Chem 278(48): 48330-48338. 6. Chu, F., et al., Protein Sci (2006) Jun 15(6) 1260-1269. 7. Peter, F., et al., (1992) J Biol Chem 267:10631-10637. 8. Allen, S. et al (2000) Blood 96(2): 560-568. 9. Sato, K et al (2001) Blood 98(6): 1852-1857. 10. Yun, S.-W. et al (2000) Brain Research Bulletin.52(5): 371-378. 11. Choukhi, A., et al (1998) J. Virol. 72:3851-3858. 12. Hoshino, T., et al (1998) Blood 91(11):4379-4386. 13. Riera, M. et a (1999) Mol. Cell Biochem. 191: 97-104. 14. Gusarova, V. et al (2001) J. Biol. Chem. 276(27): 24891-24900.

Additional Info

Detects a 98kD protein corresponding to the molecular mass of Grp94 on SDS PAGE immunoblots. Does not detect human Hsp90, Grp74 or GrpE from E. coli. Species Crossreactivity: Human, mouse, rat, bovine, canine, chicken, guinea pig, hamster, equine, monkey, porcine, rabbit, sheep, Xenopus.