Nori Human Annexin V ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of Annexin V in human. 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 ANXA5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ANXA5 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for ANXA5 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 ANXA5 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for annexin V: Annexin A5, ANXA5
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Human Annexin V ELISA Kit Summary
Alternative names for Annexin V: Annexin A5, ANXA5, ANX5, ENX2, PP4
| 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 | P08758 |
| 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 | 60 pg/mL |
| Detection Range | 0.313-20 ng/mL |
| Specificity | Human ANXA5 |
| 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:
Annexin A5 (or annexin V) is a cellular protein in the annexin group.[1] In flow cytometry, annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane. The function of the protein is unknown; however, annexin A5 has been proposed to play a role in the inhibition of blood coagulation by competing for phosphatidylserine binding sites with prothrombin and also to inhibit the activity of phospholipase A1. These properties have been found by in vitro experiments. Antibodies directed against annexin A5 are found in patients with a disease called the antiphospholipid syndrome (APS), a thrombophilic disease associated with autoantibodies against phospholipid compounds. Annexin A5 forms a shield around negatively charged phospholipid molecules. The formation of an annexin A5 shield blocks the entry of phospholipids into coagulation (clotting) reactions. In the antiphospholipid antibody syndrome, the formation of the shield is disrupted by antibodies. Without the shield, there is an increased quantity of phospholipid molecules on cell membranes, speeding up coagulation reactions and causing the blood-clotting characteristic of the antiphospholipid antibody syndrome. Annexin A5 showed upregulation in papillary thyroid carcinoma.[2] Alpha v beta 5 integrin-dependent programmed cell death was triggered by a peptide mimic of annexin V.[3] Annexin A5 as a novel player in prevention of atherothrombosis in SLE.[4] Annexin V inhibits protein kinase C.[5]
References
- Huber R, et al. (1990). The EMBO Journal. 9 (12): 3867–74.
- Sofiadis A, et al. (2012). European Journal of Endocrinology 166 (4): 657–67.
- Cardó-Vila M, et al. (2003). Molecular Cell. 11 (5): 1151–62.
- Cederholm A, Frostegård J (2007). Annals of the New York Academy of Sciences. 1108 (1): 96–103.
- Schlaepfer DD, et al. (1992). Biochemistry. 31 (6): 1886–91.
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