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Boster's GRP78 ELISA is a sandwich ELISA that can be used for the quantification of Rat GRP78 in Tissue Lysates. The ELISA utilizes an Anti-Rat GRP78 monoclonal antibody-coated plate and a Biotinylated antibody for detection which allows for an assay range of 0 to 50 ng/mL, with a sensitivity of 0.15 ng/mL. The other highlights of this kit are a quick incubation time, stable reagents, and an easy to use protocol.
| Product Name | Rat GRP78 ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7117 |
| Description | Colorimetric detection of GRP78 in rat lysates. 96wells/kit, with removable strips. |
| Cite This Product | Rat GRP78 ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7117) |
| Validated Species | Rat |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. **For positive and negative control design, consult "Tissue specificity" under Protein Target Info. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 0.15 ng/ml *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 0 - 50 ng/ml *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Tissue *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store at 4°C. |
| Description | Quantity |
| Anti-Rat GRP78 Immunoassay Plate | 1 Plate |
| Recombinant Rat GRP78 Standard | 2 vials/ 56.7ng |
| Standard and Sample Diluent | 1 vial/ 50 ml |
| Anti-Rat Grp78 Biotinylated Antibody Concentrate | 1 vial/150?L |
| 10X Wash Buffer Concentrate | 1 vial/100 ml |
| Anti-Rat GRP78 Biotinylated Antibody Diluent | 1 vial/ 13 ml |
| Streptavidin Poly HRP Concentrate | 1 vial/150 μl |
| Streptavidin Poly HRP Diluent | 1 vial/ 13 ml |
| TMB Substrate | 1 vial/ 13 ml |
| Stop Solution | 1 vial/ 13 ml |
| Plate Cover | 2 |
1. A plate reader capable of measuring absorbance at 450 nm.
2. Adjustable pipettes and a repeat pipettor.
3. Deionized or distilled water
4. Materials used for Sample Preparation
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | Hspa5 |
|---|---|
| Protein Name | Endoplasmic reticulum chaperone BiP |
| Protein Function | Endoplasmic reticulum chaperone that plays a key role in protein folding and quality control in the endoplasmic reticulum lumen (By similarity). Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10/ERdj5, probably to facilitate the release of DNAJC10/ERdj5 from its substrate (By similarity). Acts as a key repressor of the ERN1/IRE1-mediated unfolded protein response (UPR). In the unstressed endoplasmic reticulum, recruited by DNAJB9/ERdj4 to the luminal region of ERN1/IRE1, leading to disrupt the dimerization of ERN1/IRE1, thereby inactivating ERN1/IRE1. Accumulation of misfolded protein in the endoplasmic reticulum causes release of HSPA5/BiP from ERN1/IRE1, allowing homodimerization and subsequent activation of ERN1/IRE1 (By similarity). |
| Subcellular Localization | Endoplasmic reticulum lumen. |
| Uniprot ID | P06761 |
| Alternative Names | Endoplasmic reticulum chaperone BiP |
| Research Areas | Cancer, Heat Shock, Cell Signaling, Chaperone Proteins, Organelle Markers, Protein Trafficking| |
*if product is indicated to react with multiple species, protein info is based on the human gene.
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. GRP78 is also critical for maintenance of cell homeostasis and the prevention of apoptosis. Luo et al. have provided findings that suggest GRP78 is essential for embryonic cell growth and pluripotent cell survival. 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, and its protein levels are reduced in the brains of Alzheimer’s patients. Also, the induction of the GRP78 protein that results in severe glucose and oxygen deprivation could possible lead to drug resistance to anti?tumor drugs.
Click the images to enlarge.
Typical Standard Curve for the Rat GRP78 ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7117 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0 – 50 ng/ml.
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%.
1. Prepare Standard and samples in Standard and Sample Diluent.
2. Add 100 ?L 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 ?L 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 ?L 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 ?L 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 ?L of Stop Solution to each well.
14. Measure absorbance on a plate reader at 450 nm.
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