DIABLO ELISA Kit

Catalog No. ABIN1889324

This Colorimetric ELISA kit is designed for the quantitative measurement of Human DIABLO.

Quick Overview for DIABLO ELISA Kit (ABIN1889324)

Target: DIABLO (Second Mitochondria-Derived Activator of Caspase (DIABLO))

Binding Specificity: AA 56-239

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2 pg/mL - 2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma (EDTA), Plasma (heparin), Serum

Product Details DIABLO ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: Sandwich Human Diablo/SMAC ELISA Kit to quantitate Human DIABLO in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: A56-D239

Capture antibody and Detection antibody: monoclonal antibody from mouse, polyclonal antibody from goat

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: < 10 pg/mL

Components:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Material not included: Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional). Pipettes and pipette tips capable of precisely dispensing 0.5 μL through 1 mL volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500 mL graduated cylinders. Test tubes for dilution.

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Sample Volume: 100 μL

Plate: Pre-coated

Protocol: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the target amount in sample captured in plate.

Assay Precision: Intra-Assay Precision (CV%): 7.5%, 7.7%, 4.2%
Inter-Assay Precision (CV%): 8.7%, 8.8%, 5.2%

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Expiry Date: 12 months

Target Details for DIABLO

Target: DIABLO (Second Mitochondria-Derived Activator of Caspase (DIABLO))

Alternative Name: DIABLO

Background:

Background: Diablo homolog, mitochondrial, also known as second mitochondria-derived activator of caspases (SMAC), is a protein that in humans is encoded by the DIABLO gene ( IAP binding protein with low pI). It is mapped to 12q24.31. DIABLO can bind mammalian IAP homolog A (MIHA, or API3) and can also interact with MIHB, MIHC, and OpIAP, the baculoviral IAP. This gene encodes an inhibitor of apoptosis protein (IAP)-binding protein. The encoded mitochondrial protein enters the cytosol when cells undergo apoptosis, and allows activation of caspases by binding to inhibitor of apoptosis proteins. Over expression of the encoded protein sensitizes tumor cells to apoptosis. A mutation in this gene is associated with young-adult onset of nonsyndromic deafness-64.

Gene Full Name: diablo IAP-binding mitochondrial protein

Gene ID: 56616

UniProt: Q9NR28

Pathways: Apoptosis, Caspase Cascade in Apoptosis, Positive Regulation of Endopeptidase Activity