Diablo Homolog (Drosophila) (DIABLO) antibody

Antigen: Diablo Homolog (Drosophila) (DIABLO)

Synonyms: LOC100074947, LOC100022080, Smac, AU040403, 0610041G12Rik, 1700006L01Rik, MGC105680, SMAC, SMAC3, DIABLO-S, FLJ10537, FLJ25049, smac, smac3, diablo-s, DIABLO, DBO, MGC134245, DKFZp469M2435

Clonality: Polyclonal

Host: Rabbit

Reactivity: Cow (Bovine), Human, Mouse (Murine), Rat (Rattus), Sheep (Ovine)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN106863

Additional Information

Alternative name: Smac/DIABLO

Immunogen: This whole rabbit serum was prepared by repeatedimmunizations with recombinant His6-tagged human Smac/DIABLOprotein (amino acids 56-239).

Format: Liquid (sterile filtered)

Description: Apoptosis is a conserved cell suicideprogram essential for the development and homeostasis of multi-cellular organisms. Abnormal inhibition of apoptosis is a hallmark ofcancer and autoimmune diseases, whereas excessive cell death isfound in neurodegenerative disorders such as Alzheimers disease.Executioners of the apoptotic program are cysteine proteasestermed caspases that exist as inactive zymogens in living cells andare activated during apoptosis. Active caspases cleave keyintracellular protein substrates, resulting in the characteristicmorphological changes associated with apoptosis. The release ofcytochrome c from the mitochondria triggers the oligomerization ofApaf-1 in an ATP/dATP-dependent manner and induces theautoactivation of caspase-9. Active caspase-9 in turn activatesdownstream effector caspases including caspase -3, -6 and -7. Given the potentially irreversible cascade triggered by theactivation of initiator caspases, it is crucial that activation ofcaspases be tightly regulated. A number of cellular proteins havebeen shown to modulate caspase activation and activity. Theseinclude members of the Inhibitor of the Apoptosis Protein (IAP)family, such as XIAP, c-IAP-1 and c-IAP-2, which block celldeath by inhibiting the activity of the effector caspases -3 and -7and the initiator caspase-9. Smac/DIABLO (SecondMitochondria-derived Activator of Caspase) is a mitochondrialprotein of 27 kDa that is released from mitochondria duringapoptosis. Smac/DIABLO promotes caspase activation throughbinding and neutralizing IAPs.

Specificity: This antiserum is directed against humanSmac/DIABLO and is useful in determining its presence in variousassays.

Application Details

Application Notes: Recommended Dilutions: This product was assayed by immunoblotand was found to be reactive at a 1:1000 to 1:2000 dilution. A 27 kDaband corresponding to human Smac/DIABLO protein is detected.Detection occurs using HRP Donkey anti-Rabbit IgG and ECL. As littleas 10 ng of recombinant protein can be detected by ECL. Theresearcher should determine optimal titers for other applications. Note: This antibody reacts with human Smac/DIABLO protein by immunoblot. Lysates fromhuman HeLa and LNCaP cells are positive for Smac/DIABLO. Other animal tissues have not been tested.

Concentration: 85 mg/ml (by Refractometry)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C. For extended storage aliquot contents and freeze at -20° C or below.Avoid cycles of freezing and thawing. Dilute only prior to immediate use. Expiration date is one (1) year from dateof restoration.

Research Area: Apoptosis/Necrosis

Restrictions: For Research Use only

Images

Anti-Smac is shown to detect a 25-26 kDa band in partially purified recombinant human Smac protein by immunoblot. Lanes 1-3 are loaded with 1, 10 and 100 ng of protein per lane, respectively. The blot was incubated overnight with a 1:1000 dilution of anti-Smac in TBST. Detection occurs using a 1:1000 dilution of HRP Goat-a-Rabbit with visualization via ECL. Film exposure approximately 1

Immunoblot using anti-Smac detects a 26 kDa band when 1 ug of recombinant Smac is applied (lane 1). Lane 2 shows Smac detection when 30 ug of 1% NP-40 treated cell lysate from HeLa cells is applied. Lanes 3 & 4 show 30 ug each of cytosolic fractions from HeLa cell lysates both with (lane 3) and without (lane 4) treatment with 30 uM etoposide. Recombinant Smac migrates slower than the native form because of the His6-tag. The blot was incubated overnight with a 1:1000 dilution of anti-Smac in TBST. Detection occurs using a 1:1000 dilution of HRP Goat-a-Rabbit with visualization via ECL. Film exposure approximately 1