Human Diablo/SMAC ELISA Kit EZ-SetTM (DIY Antibody Pairs)

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.

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.

Cancer, Cell Biology, Cell Death, Invasion/Microenvironment, Metabolism, Metabolism Processes, Mitochondrial, Mitochondrial Markers, Mitochondrial Metabolism, Pathways and Processes, Receptors

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% 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% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 112 545 1134 129 577 1133 Mean (pg/ml) 112 545 1134 129 577 1133 Standard deviation 6.27 23.98 58.97 7.99 40.97 77.04 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% CV (%) 5.6% 4.4% 5.2% 6.2% 7.1% 6.8%

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.

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/tubex3 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. 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/tubex3 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. Catalog number Description Quantity Storage of opened/reconstituted material 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-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) 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) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0838-ST Lyophilized recombinant human DIABLO standard 10 ng/tubex3 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. EK0838-ST Lyophilized recombinant human DIABLO standard 10 ng/tubex3 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.

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.)

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 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 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 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 O.D. 0.024 0.049 0.096 0.193 0.358 0.773 1.338 2.294

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. 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)