Nori Bovine HSP70 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of HSP70 in bovine. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for HSP70 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HSP70 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for HSP70 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of HSP70 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for HSP70: Heat shock protein 70, HSPA1A, HSP70-1
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Bovine HSP60 ELISA Kit Summary
Alternative names for HSP70: Heat shock protein 70, HSPA1A, HSP70-1
Alternative names for bovine: Cattle, cow, bull, buffalo
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | Q27975 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 30 pg/mL |
| Detection Range | 0.156-10 ng/mL |
| Specificity | Bovine HSP70 |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
The 70 kilodalton heat shock proteins (HSP70s or DnaK) are a family of conserved ubiquitously expressed heat shock proteins. Proteins with similar structure exist in virtually all living organisms. The Hsp70s are an important part of the cell’s machinery for protein folding, and help protect cells from stress. The Hsp70 system interacts with extended peptide segments of proteins as well as partially folded proteins to cause aggregation of proteins in key pathways to deregulate activity.[1] HOP (the Hsp70/Hsp90 Organizing Protein) can bind to both Hsp70 and Hsp90 at the same time, and mediates the transfer of peptides from Hsp70 to Hsp90.[2] Hsp70 can be phosphorylated[3] which regulates several of its functions.[4][5][6] Hsp70 proteins can protect cells from thermal or oxidative stress. Hsp70 seems to be able to participate in disposal of damaged or defective proteins. Interaction with CHIP–an E3 ubiquitin ligase–allows Hsp70 to pass proteins to the cell’s ubiquitination and proteolysis pathways.[7] HSP70 can improve overall protein integrity and directly inhibits apoptosis.[8] Hsp70 blocks the recruitment of procaspase-9 to the Apaf-1/dATP/cytochrome c apoptosome complex. Hsp70 is shown to interact with Endoplasmic reticulum stress sensor protein IRE1alpha thereby protecting the cells from ER stress – induced apoptosis. Hsp70 not only saves important components of the cell but also directly saves the cell as a whole. Hsp70 is overexpressed in malignant melanoma[9] and under expressed in renal cell cancer.[10]
References
- Mashaghi A, et al. (2016). Nature. 539 (7629): 448–451.
- Wegele H, et al. (2004). Rev. Physiol. Biochem. Pharmacol. 151. pp. 1–44.
- Cvoro A, et al. (1999). Cell Biology International. 23 (4): 313–20.
- Gao T, Newton AC (2002). The Journal of Biological Chemistry. 277 (35): 31585–92.
- Truman AW, et al. (2012). Cell. 151 (6): 1308–18.
- Muller P, et al. (2013). Oncogene. 32 (25): 3101–10.
- Lüders J, et al. (2000). The Journal of Biological Chemistry. 275 (7): 4613–7.
- Beere HM, et al. (2000). Nature Cell Biology. 2 (8): 469–75.
- Ricaniadis N, et al. (2001). European Journal of Surgical Oncology. 27 (1): 88–93.
- Ramp U, et al. (2007). Histology and Histopathology. 22 (10): 1099–107.
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