Nori Equine Granzyme A ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of Granzyme A in equine. 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 GZMA has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any GZMA present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for GZMA 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 GZMA bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for granzyme A: GZMA, CTLA3
This product is for laboratory research use only, not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Equine Granzyme A ELISA Kit Summary
Alternative names for Granzyme A: GZMA, CTLA3
Alternative names for equine: Horse
| 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 | F6TIE2 |
| 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 | 6 pg/mL |
| Detection Range | 31.25-2000 pg/mL |
| Specificity | Equine Granzyme A |
| 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:
Granzyme A (GZMA) is a cytotoxic T-lymphocyte serine protease that is encoded by the GZMA gene, and is one of the five granzymes.[1][2] Cytolytic T lymphocytes (CTL) and natural killer (NK) cells share the remarkable ability to recognize, bind, and lyse specific target cells. They are thought to protect their host by lysing cells bearing on their surface ‘nonself’ antigens, usually peptides or proteins resulting from infection by intracellular pathogens. Granzyme A is a T cell- and natural killer cell-specific serine protease that may function as a common component necessary for lysis of target cells by cytotoxic T lymphocytes and natural killer cells. GZMA from cytotoxic lymphocytes cleaves and activates GSDMB to induce target cell pyroptosis.[3] Granzyme A in human platelets regulates the synthesis of proinflammatory cytokines by monocytes in aging.[4] GzmA produced by lesional CD8 T cells has the capacity to specifically increase chemokine secretion from inflamed keratinocytes, thereby sustaining the focal inflammation in psoriasis lesions by amplifying a chemotactic inflammatory loop.[5] GzmA plays an unfavorable role in host defense during pneumococcal pneumonia by a mechanism that does not depend on natural killer cells.[6] Granzyme A binds to and cleaves nucleolin in vitro.[7] Granzyme A activates an endoplasmic reticulum-associated caspase-independent nuclease to induce single-stranded DNA nicks.[8] Granzyme A mediates cell death by enhancing cleavage of the oxidative repair protein Ape1.[9]
References
- Masson D, et al. (1986). FEBS Lett. 208 (1): 84–8.
- Gershenfeld HK, et al. (1988). Proc. Natl. Acad. Sci. U.S.A. 85 (4): 1184–8.
- Zhou Z, et al. (2020) Science 368 (6494).
4. Campbell RA, (2018) J. Immunol. 200 (1), 295-304.
5. Cheuk S, et al. (2017) Exp. Dermatol. 26 (9), 824-827.
- van den Boogaard FE, et al. (2016) Am. J. Physiol. Lung Cell Mol. Physiol. 311 (2), L507-L516.
- Pasternack MS, et al. (1991) Biol. Chem. 266 (22), 14703-14708.
- Beresford PJ, et al. (2001). J. Biol. Chem. 276 (46): 43285–93.
- Fan Z, et al. (2003). Nat. Immunol. 4 (2): 145–53.
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