Human IRE1 alpha - Ready-To-Use ELISA Kit (Colorimetric) Summary
| Description |
The Ready-To-Use ELISA kit offers pre-diluted detection reagents and a shorter experimental time. Assay Length: 3 hours |
| Standard Curve Range |
0.156 - 10 ng/mL (example only; lot dependent) |
| Sensitivity |
0.071 ng/mL (example only; lot dependent) |
| Assay Type |
Sandwich ELISA |
| Inter-Assay |
%CV < 12 (example only; lot dependent) |
| Intra-Assay |
%CV < 10 (example only; lot dependent) |
| Sample Volume |
100 uL |
| Kit Type |
ELISA Kit (Colorimetric) |
| Gene |
ERN1 |
Applications/Dilutions
Packaging, Storage & Formulations
| Storage |
Storage of components varies. See protocol for specific instructions. |
Kit Components
|
Components
|
- Detection Solution A
- Detection Solution B
- Instruction manual
- Plate sealer for 96 wells
- Pre-coated 96T strip plate
- Standard Diluent
- Standard
- Stop Solution
- TMB Substrate
- Wash Buffer (30 x concentrate)
|
Alternate Names for Human IRE1 alpha - Ready-To-Use ELISA Kit (Colorimetric)
Background
Accumulation of misfolded proteins in the endoplasmic reticulum (ER) activates the unfolded protein response (UPR) and upregulates ER molecular chaperones in order to cope with ER stress. UPR is initiated by three ER-localized protein sensors: PERK (PKR-like ER kinase), ATF (activating transcription factor 6), and IRE1 alpha (inositol-requiring enzyme 1 alpha) (1). IRE1 alpha is correlated with X-box binding protein (XBP1) as a potent UPR transcriptional activator (2).
Transcriptional activation of UPR-responsive genes are regulated by the ATF6 and IRE1-XBP1 pathways, which are often regulated by HIFs and contribute to cell survival under ER hypoxic stress (3). UPR signaling can a) inhibit protein translation to restore cell function, b) activate signaling to increase production of molecular chaperones for protein folding, and c) initiate ubiquitination signaling that leads to degradation of misfolded proteins in ER.
IRE1 alpha acts as the sensor of unfolded proteins in the ER. IRE1 alpha not only promotes cell survival but can initiate apoptosis when accumulation of unfolded proteins in the ER causes stress. IRE1 alpha is essential for viability under stress conditions that cause unfolded proteins to accumulate in the ER. IRE1 alpha is a transmembrane protein that has both serine-threonine kinase and endoribonuclease activities and has a theoretical molecular weight of 110 kDa. When detecting phospho-IRE1 alpha, it is recommended to normalize its band intensity with total IRE1 alpha.
References
1. Zheng, W., Xie, W., Yin, D., Luo, R., Liu, M., & Guo, F. (2019). ATG5 and ATG7 induced autophagy interplays with UPR via PERK signaling. Cell Commun Signal, 17(1), 42. doi:10.1186/s12964-019-0353-3
2. Cho, Y. M., Kim, D. H., Lee, K. H., Jeong, S. W., & Kwon, O. J. (2018). The IRE1alpha-XBP1s pathway promotes insulin-stimulated glucose uptake in adipocytes by increasing PPARgamma activity. Exp Mol Med, 50(8), 102. doi:10.1038/s12276-018-0131-0
3. Xia, Z., Wu, S., Wei, X., Liao, Y., Yi, P., Liu, Y., . . . Liu, J. (2019). Hypoxic ER stress suppresses beta-catenin expression and promotes cooperation between the transcription factors XBP1 and HIF1alpha for cell survival. J Biol Chem, 294(37), 13811-13821. doi:10.1074/jbc.RA119.008353
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are
guaranteed for 6 months from date of receipt.
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Product General Protocols
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FAQs for IRE1 alpha ELISA Kit (NBP3-31953). (Showing 1 - 1 of 1 FAQ).
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What is the difference between the Quantikine and Duoset ELISA kits?
- Our Quantikine kits are fully optimized and validated for the sample types listed on the product specific webpage and datasheet, as this can vary between kits. Each kit is supplied ready to use with one pre-coated 96-well plate, a detection antibody directly conjugated to HRP, and all other necessary reagents. These kits are ideal for researchers who want the convenience of a ready to use and optimized ELISA product. Some of these kits are also available prepackaged in larger 6 and 50 plate sizes.Our DuoSet Kits, in contrast, are ELISA development kits containing the capture and detection antibody, the mass-value calibrated standard, and streptavidin-HRP to prepare approximately 5 or 15 plates. Ancillary reagents will need to be used/purchased, and for most kits, we will recommend one of our Ancillary Reagent Kits, which contain the reagents we use ourselves in-house. DuoSet kits are validated only for cell culture supernatant samples and therefore require further development and validation for accurate measurement in more complex samples such as serum and plasma. Our DuoSet Kits offer an economical, flexible alternative for the experienced ELISA user.