DIABLO ELISA Kit

Produktnummer ABIN1889324

Dieses Colorimetric ELISA-Kit wurde entwickelt für die quantitative Messung von Human DIABLO.

Kurzübersicht für DIABLO ELISA Kit (ABIN1889324)

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

Bindungsspezifität: AA 56-239

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 31.2-2000 pg/mL

Applikation: ELISA

Proben: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Produktdetails DIABLO ELISA Kit

Untere Nachweisgrenze: 31.2 pg/mL

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

Marke: PicoKine™

Analytische Methode: Quantitative

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

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

Sensitivität: <10pg/mL

Benötigtes Material: 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

Anwendungsinformationen

Applikationshinweise: 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.

Kommentare:

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. .

Plattentyp: Pre-coated

Protokoll: 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.

Testdurchführung:

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.

Testpräzision:

• 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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid multiple freeze-thaw cycles.

Lagerung: -20 °C,4 °C

Informationen zur Lagerung: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Haltbarkeit: 12 months

Details zu DIABLO

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

Andere Bezeichnung: DIABLO

Hintergrund:

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.

Gen-ID: 56616

UniProt: Q9NR28

Pathways: Apoptose, Caspase Kaskade in der Apoptose, Positive Regulation of Endopeptidase Activity