Technical Data
G3057-13
Glucose Regulated Protein 75 (Grp75)
Description:
In response to adverse changes in their environment, cells from many organisms increase the expression of a class of proteins referred to as heat shock or stress proteins. One class of stress proteins, termed the Hsp70 family, is comprised of multiple members, all of which bind ATP in vitro, but which are localized within different intracellular compartments. These include: i) Hsc70 (or constitutive form) present within the cytosol/nucleus; ii) Hsp70 (inducible form) present within the cytosol/nucleus/nucleolus; iii) the constitutive glucose-regulated 78kD (or BiP) protein present within the lumen of the endoplasmic reticulum; and iv) the constitutive glucose regulated 75kD protein present within the mitochondrial matrix. Members of the Hsp70 family are thought to funtion as molecular chaperones (2), assisting in the folding of other proteins in various intracellular compartments (3). Grp75 is localized in the mitochondrial matrix, where, in concert with Hsp60, is thought to participate in the re-folding of proteins translocated into this organelle (4). Like its E. coli homolog DnaK (5), Grp75 possesses a cation-dependent ATPase activity (6,7) thought to be central to its function as a chaperone.

Applications:
Suitable for use in Western Blot and Immunohistochemistry. Other applications have not been tested.

Recommended Dilution:
Western Blot (Colorimetric): 1:1000. Sufficient for detection of Grp75 in 20ug of HeLa (heat shocked) cell lysate by colorimetric immunoblot analysis using goat anti-mouse IgG:AP as the secondary antibody.
Immunohistochemistry: 1:50-1:200 using human breast cancer tissue.
Optimal dilutions to be determined by researcher.

Positive Controls:
HeLa Heat Shocked Cell Lysate

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. 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.
TypeIsotypeCloneGrade
MabIgG14E86Purified
SizeStorageShippingSourceHost
50ul-20CBlue IceHumanMouse
Concentration:
Not determined
Immunogen:
Grp75 isolated from human Molt-4 cells
Purity:
Purified by ammonium sulfate precipitation.
Form
Supplied as a liquid in PBS, pH 7.2, 50% glycerol.
Specificity:
Detects a ~75kD protein, corresponding to the apparent molecular mass of Grp75 on SDS-PAGE. Species Crossreactivity: Human, monkey, mouse, rat, bovine, canine, chicken, Drosophila, guinea pig, hamster, porcine, rabbit, sheep and Xenopus.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Celis, J.; Electrophoresis 11:989-1071 (1990). 2. Ellis, R.J.; Semin. Cell Biol. 1:1-9 (1990). 3. Gething, M.-J., and Sambrook, J.; Nature 355:33-45 (1992). 4. Manning-Krieg, U.C., Scherer, P.E., and Schatz, G.; EMBO J. 10:3273-3280 (1991). 5. Liberek, K., Skowya, D., Zylicz, M., Johnson, C., and Georgopoulos, C.; J. Biol. Chem., 266: 14491-14496 (1991). 6. Mizzen, L.A., Kabiling, A.N., and Welch, W.J.; Cell Regulation 2:165-179 (1991). 7. Leustek, U.K., Dalie, B., Amir-Shapira, D., Brot, N., and Weissbach, H.; PNAS USA
86:7805-7808 (1989). 8. Plakidou-Dymock, S., McGivan, J.D.; Biochem. Biophys. Acta 1224:189-197 (1994). 9. Chen, J., Graham, S.H., Zhu, R.L., Simon, R.P.C. J. Cerebral Blood Flow & Metabolism 16:566-577 (1996). 10. He, D.; Hagen, S.J.; Pothoulakis, C.; Chen, M.; Medina, N.D.; Warny, M.; LaMont, J.T.; Gastroenterology 119(1):139-150 (2000). 11. Milewski, M.I., Mickle, J.E., Forrest, J.K., Stanton, B.A., Cutting, G.R.; J Biol Chem 277(37):34462-34470 (2002). 12. Ilieva, H., Nagano, I., Murakami, T., Shiote, M., Manabe, Y., Abe, K.; Neurosci Lett 332(1):53-56 (2002). 13. Mitchell, C.R., Harris, M.B., Cordaro, A.R., Starnes, J.W.; J Appl Physiol 93(2):526-530 (2002).