Smac/Diablo Antibody [J14F1] | Primary Antibodies

Application: Reactivity: Human, Monkey

Lane 1: 293, Lane 2: Hela, Lane 3: Jurkat

Usage Information

Dilution
WB1:1000 IP1:100 IHC1:400

Application
WB, IP, IHC

Reactivity
Human, Monkey

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
21 kDa

Positive Control	Human breast carcinoma; Human lung carcinoma; Human colon carcinoma; Human renal cell carcinoma; 293 cells; HeLa cells; Jurkat cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
Smac/Diablo Antibody [J14F1] detects endogenous levels of total Smac/Diablo protein.

Clone
J14F1

Synonym(s)
Diablo IAP-binding mitochondrial protein; Diablo IAP-binding mitochondrial protein; Direct IAP-binding protein with low pI; Second mitochondria-derived activator of caspases (SMAC); DIABLO; SMAC

Background
Smac/DIABLO (Second mitochondria-derived activator of caspases/Direct IAP-binding protein with low pI) is a dimeric protein synthesized as a 239-residue mitochondrial precursor that undergoes IMP complex-mediated cleavage of its N-terminal targeting sequence, generating the mature form with an exposed AVPI tetrapeptide motif (Ala67-Val68-Pro69-Ile70). It adopts a compact α-helical fold, with three long helices forming a flat, saddle-shaped dimer interface and a hydrophobic cleft that engages IAP BIR2/BIR3 domains through N-terminal IBM (IAP-binding motif) docking, supplemented by dimer interface contacts. Upon apoptotic stimuli such as DNA damage or growth factor withdrawal, Bax/Bak oligomerization permeabilizes the outer mitochondrial membrane, releasing mature Smac/DIABLO into the cytosol. Here, its AVPI motif binds XIAP BIR3 (releasing caspase-9) and the BIR2-linker (releasing caspase-3/7), while homodimerization enhances affinity for cIAP1/2 via BIR1-BIR3 interfaces, triggering their autoubiquitination/degradation and non-canonical NF-κB activation. This cascade relieves IAP-mediated caspase suppression, amplifying the executioner phase of apoptosis, while Smac’s C-terminal region further interacts with SMACOV/USP9X for IAP stabilization feedback. Smac/DIABLO downregulation is linked to chemoresistance and poor prognosis in various cancers (lung, colon, leukemia), while germline mutations impair apoptosis and contribute to autoimmune lymphoproliferation.

Background

Smac/DIABLO (Second mitochondria-derived activator of caspases/Direct IAP-binding protein with low pI) is a dimeric protein synthesized as a 239-residue mitochondrial precursor that undergoes IMP complex-mediated cleavage of its N-terminal targeting sequence, generating the mature form with an exposed AVPI tetrapeptide motif (Ala67-Val68-Pro69-Ile70). It adopts a compact α-helical fold, with three long helices forming a flat, saddle-shaped dimer interface and a hydrophobic cleft that engages IAP BIR2/BIR3 domains through N-terminal IBM (IAP-binding motif) docking, supplemented by dimer interface contacts. Upon apoptotic stimuli such as DNA damage or growth factor withdrawal, Bax/Bak oligomerization permeabilizes the outer mitochondrial membrane, releasing mature Smac/DIABLO into the cytosol. Here, its AVPI motif binds XIAP BIR3 (releasing caspase-9) and the BIR2-linker (releasing caspase-3/7), while homodimerization enhances affinity for cIAP1/2 via BIR1-BIR3 interfaces, triggering their autoubiquitination/degradation and non-canonical NF-κB activation. This cascade relieves IAP-mediated caspase suppression, amplifying the executioner phase of apoptosis, while Smac’s C-terminal region further interacts with SMACOV/USP9X for IAP stabilization feedback. Smac/DIABLO downregulation is linked to chemoresistance and poor prognosis in various cancers (lung, colon, leukemia), while germline mutations impair apoptosis and contribute to autoimmune lymphoproliferation.

References
https://pubmed.ncbi.nlm.nih.gov/11140637/ https://pubmed.ncbi.nlm.nih.gov/11140638/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%