| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | Diablo homolog, mitochondrial; IAP-binding protein with low pI; Second mitochondria-derived activator of caspase; Smac; DIABLO; SMAC |
| Assay Time | |
| Assay Type | |
| Detection Range | |
| Expression System | |
| Gene ID | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: A56-D239 |
| Product Type | |
| Reactivity | |
| Sample Type(s) | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
| Sensitivity | |
| Storage | |
| Target | |
| UniProt # |
Background
Also known as: Diablo homolog, mitochondrial, IAP-binding protein with low pI, Second mitochondria-derived activator of caspase, Smac, DIABLO.
Human Diablo/SMAC (DIABLO) is widely studied as a molecular readout in experimental models where changes in protein abundance reflect underlying biology. This target is frequently investigated in Molecular & Cellular Biology research contexts. Proteases and extracellular matrix (ECM) components are central to tissue architecture and remodeling. In many experimental contexts, changes in ECM-related proteins reflect shifts in cell adhesion, migration, barrier integrity, or matrix turnover.
Biological function and remodeling context
Matrix remodeling is influenced by the balance between synthesis and degradation, often regulated by inflammatory cues, mechanical stress, and growth-factor signaling. Protease activity can unmask or release bioactive fragments, while altered ECM composition can feed back on cell behavior through mechanotransduction and receptor engagement.
Why it matters in research
- Remodeling readout: Quantification can support studies of fibrosis, wound repair, and invasion models.
- Microenvironment state: Levels may reflect stromal activation, barrier disruption, or matrix turnover.
- Mechanistic linkage: Pairing with inflammatory and growth-factor markers can clarify drivers of remodeling.
Disease and translational relevance
ECM remodeling and protease regulation are frequently discussed in the literature across oncology, cardiovascular, pulmonary, and inflammatory disease models. Interpretation of abundance should consider whether the measured analyte represents pro-forms, active forms, or fragments, and whether binding partners in the matrix influence detectability.
Sample data
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.024 | 0.049 | 0.096 | 0.193 | 0.358 | 0.773 | 1.338 | 2.294 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 112 | 545 | 1134 | 129 | 577 | 1133 |
| Standard deviation | 6.27 | 23.98 | 58.97 | 7.99 | 40.97 | 77.04 |
| CV (%) | 5.6% | 4.4% | 5.2% | 6.2% | 7.1% | 6.8% |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0838-CA | Rabbit anti- human DIABLO polyclonal antibody (Capture Antibody) | 500 µl, 0.4 mg/mL (recommended dilution 1:100) | Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. |
| EZ0838-DA | Biotinylated goat anti- human DIABLO polyclonal antibody (Detection Antibody) | 500 µl, 10 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0838-ST | Lyophilized recombinant human DIABLO standard | 10 ng/tube×3 | Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. |
Materials required but not provided
- Microplate reader in standard size.
- Automated plate washer.
- Incubator.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- 96 well microplate (Cat# AR1100)
- Plate Sealers.
- Capture Antibody Diluent: PBS.
- Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered.
- Substrate Solution: Tetramethylbenzidine (Cat# AR1104)
- Stop Solution: 2 N H2SO4 (Cat# AR1105)
- Wash Buffer (PBS and PBS-T). Recipes:
- PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 µm filtered.
- PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4.
- *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
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