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: 31.2-2000 pg/mL

Application: ELISA

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

Product Details DIABLO ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Diablo/SMAC

Brand: PicoKine™

Analytical Method: Quantitative

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

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

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. 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. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

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

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.

Comment:

Tissue Specificity: Ubiquitously expressed with highest expression in testis. Expression is also high in heart, liver, kidney, spleen, prostate and ovary. Low in brain, lung, thymus and peripheral blood leukocytes. Isoform 3 is ubiquitously expressed. .

Plate: Pre-coated

Protocol: human Diablo ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Diablo has been precoated onto 96-well plates. Standards(E.coli, A56-D239) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Diablo 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 human Diablo amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL human Diablo standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human Diablo standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 112, Standard deviation: 6.27, CV(%): 5.6
• Sample 2: n=16, Mean(pg/ml): 545, Standard deviation: 23.98, CV(%): 4.4
• Sample 3: n=16, Mean(pg/ml): 1134, Standard deviation: 58.97, CV(%): 5.2,
• Sample 1: n=24, Mean(pg/ml): 129, Standard deviation: 7.99, CV(%): 6.2
• Sample 2: n=24, Mean(pg/ml): 577, Standard deviation: 40.97, CV(%): 7.1
• Sample 3: n=24, Mean(pg/ml): 1133, Standard deviation: 77.04, CV(%): 6.8

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

Expiry Date: 12 months

Target Details for DIABLO

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

Alternative Name: DIABLO

Background:

Protein Function: Promotes apoptosis by activating caspases in the cytochrome c/Apaf-1/caspase-9 pathway. Acts by opposing the inhibitory activity of inhibitor of apoptosis proteins (IAP). Inhibits the activity of BIRC6/bruce by inhibiting its binding to caspases. Isoform 3 attenuates the stability and apoptosis- inhibiting activity of XIAP/BIRC4 by promoting XIAP/BIRC4 ubiquitination and degradation through the ubiquitin-proteasome pathway. Isoform 3 also disrupts XIAP/BIRC4 interacting with processed caspase-9 and promotes caspase-3 activation. Isoform 1 is defective in the capacity to down-regulate the XIAP/BIRC4 abundance. .

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(direct 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.

Synonyms: Diablo homolog, mitochondrial,Direct IAP-binding protein with low pI,Second mitochondria-derived activator of caspase,Smac,DIABLO,SMAC,

Full Gene Name: Diablo homolog, mitochondrial

Cellular Localisation: Mitochondrion . Released into the cytosol when cells undergo apoptosis.

Gene ID: 56616

UniProt: Q9NR28

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