|Product Name||Rat GRP78 ELISA Kit|
Colorimetric detection of GRP78 in rat lysates
|Detection Method||Colorimetric Assay|
|Assay Type||Sandwich ELISA (Enzyme-linked Immunosorbent Assay)|
|Utility||ELISA kit used to quantitate GRP78 concentration in rat samples.|
|Assay Range||0 - 50 ng/ml|
|Precision||Intra-Assay Precision: Three samples of known concentration were assayed thirty times on one plate. The intra-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <10%.
Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays. The inter-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <15%.
|Incubation Time||30 minutes|
|Number of Samples||40 samples in duplicate|
|Other Resources||, Kit Booklet , MSDS|
|Shipping Temperature||Blue Ice|
|Product Type||ELISA Kits|
|Assay Overview||1. Prepare Standard and samples in Standard and Sample Diluent.
2. Add 100 µL of Standard or sample to appropriate wells.
3. Cover plate with Plate Sealer and incubate at 37°C for 2 hours.
4. Wash plate four times with 1X Wash Buffer.
5. Add 100 µL of Biotinylated Antibody Working Solution to each well.
6. Cover plate with Plate Sealer and incubate at at 37°C for 2 hours.
7. Wash plate four times with 1X Wash Buffer.
8. Add 100 µL of Streptavidin Poly HRP Working Solution to each well.
9. Cover plate with Plate Sealer and incubate at 37°C for 30 minutes.
10. Wash plate four times with 1X Wash Buffer.
11. Add 100 µL of TMB Substrate to each well.
12. Develop the plate in the dark at room temperature for 30 minutes.
13. Stop reaction by adding 100 µL of Stop Solution to each well.
14. Measure absorbance on a plate reader at 450 nm.
|Cite This Product||Rat GRP78 ELISA Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-137)|
|Alternative Names||Grp78 ELISA Kit, Binding immunoglobulin protein ELISA Kit, BiP ELISA Kit, 78 kDa glucose-regulated protein ELISA Kit, heat shock 70 kDa protein 5 ELISA Kit|
|Research Areas||Cancer, Heat Shock, Cell Signaling, Chaperones, Organelle Markers, Trafficking|
|Scientific Background||GRP78 is a ubiquitously expressed, 78-kDa glucose-regulated protein, and is commonly referred to as an immunoglobin chain binding protein (BiP). The BiP proteins are categorized as stress response proteins because they play an important role in the proper folding and assembly of nascent protein and in the scavenging of misfolded proteins in the endoplasmic reticulum lumen. Translation of BiP is directed by an internal ribosomal entry site (IRES) in the 5’ nontranslated region of the BiP mRNA. BiP IRES activity increases when cells are heat stressed (1).
GRP78 is also critical for maintenance of cell homeostasis and the prevention of apoptosis (2). Luo et al. have provided findings that suggest GRP78 is essential for embryonic cell growth and pluripotent cell survival (3).
In terms of diseases, GRP78 has been shown to be a reliable biomarker of hypoglycemia, to serve a neuroprotective function in neurons exposed to glutamate and oxidative stress (4), and its protein levels are reduced in the brains of Alzheimer’s patients (5). Also, the induction of the GRP78 protein that results in severe glucose and oxygen deprivation could possible lead to drug resistance to antitumor drugs (6, 7).
|References||1. Cho S., et al. (2007). Mol Cell Biol 27(1): 368-83.
2. Yang Y., et al. (1998) J Biol Chem 273: 25552-25555.
3. Luo S., et al (2006) 26 (15): 5688-97.
4. Yu Z., et al. (1999) Exp Neurol. 15: 302-314.
5. Koomagi R., et al. (1999) Anticancer Res. 19:4333- 4336.
6. Laquerre S., et al. (1998) J. Virology 72: 4940-4949.
7. Dong D., et al. (2005) Cancer Res 65(13): 5785-91.
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