Anti-Smac (mouse CT) Antibody (1411)
| Host | Quantity | Applications | Species Reactivity | Data Sheet | |
|---|---|---|---|---|---|
| Rabbit | 100ug | ELISA,WB,IHC-P,IF,IP | Human, Mouse, Rat |  |
Overview
Product Name Anti-Smac (mouse CT) Antibody (1411)
Description Anti-mSmac (CT) Rabbit Polyclonal Antibody
Target Smac (mouse CT)
Species Reactivity Human, Mouse, Rat
Applications ELISA,WB,IHC-P,IF,IP
Host Rabbit
Clonality Polyclonal
Isotype IgG
Immunogen Synthetic peptide corresponding to aa 222-237 of mouse Smac (accession no. AF203914).
Properties
Form Liquid
Concentration Lot Specific
Formulation PBS, pH 7.4.
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Format Purified
Purification Purified by peptide immuno-affinity chromatography
Specificity Information
Specificity This antibody recognizes human, mouse, and rat Smac (25kDa).
Target Name Diablo IAP-binding mitochondrial protein
Target ID Smac (mouse CT)
Uniprot ID Q9JIQ3
Alternative Names Diablo homolog, mitochondrial, Direct IAP-binding protein with low pI, Second mitochondria-derived activator of caspase, Smac
Gene Name Diablo
Gene ID 66593
Accession Number NP_075721
Sequence Location Mitochondrion
Biological 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 (By similarity). {PubMed:10929712}.
Research Areas Apoptosis
Background Inhibitors of apoptosis proteins (IAPs) regulate programmed cell death by inhibiting members of the caspase family of enzymes. A novel mammalian protein that binds to IAPs and neutralizes the inhibitory effect of IAPs on caspases has been identified and designated Smac/DIABLO. Smac is a mitochondrial protein that is released along with cytochrome c during apoptosis and activates the cytochrome c/Apaf- 1/caspase-9 pathway. The N-terminal amino acids of Smac are required for binding to IAPs and for activation of caspases. Smac is expressed in a variety of human and mouse tissues.
Application Images
Description Western Blot Validation in (A and B) Mouse Heart Tissue Lysate and (C) Rat Heart Tissue Lysate
Loading: 15 ug of lysates per lane. Antibodies: Smac 1411 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.A Mouse heartB Mouse heart and blocking peptideC Rat heart
Loading: 15 ug of lysates per lane. Antibodies: Smac 1411 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.A Mouse heartB Mouse heart and blocking peptideC Rat heart
Description Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: Smac 1409 (1 ug/mL), Smac 1411 (1 ug/mL), and beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: Smac 1409 (1 ug/mL), Smac 1411 (1 ug/mL), and beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Western Blot Validation in Human, Mouse and Rat Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: Smac 1411 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: Smac 1411 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Immunofluorescence Validation of Smac in Mouse Spleen Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Spleen Cells labeling Smac with 1411 at 10 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Immunofluorescent analysis of 4% paraformaldehyde-fixed Mouse Spleen Cells labeling Smac with 1411 at 10 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Description Immunohistochemistry Validation of Smac in Mouse Spleen Tissue
Immunohistochemical analysis of paraffin-embedded Mouse Spleen Tissue using anti-Smac antibody (1411) at 2 ug/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Immunohistochemical analysis of paraffin-embedded Mouse Spleen Tissue using anti-Smac antibody (1411) at 2 ug/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Description KO Validation in Mouse Fibroblasts and Myoblasts (Ho et al., 2416)
The indicated MEFs or MEMs were exposed to 2 uM STS for 4 h and analyzed by Western blot. Accumulation of Smac/Diablo in mitochondrion-depleted cytosol fractions fromSTS-treated Apaf-1 KO cells were detected by anti-smac antibodies. Smac expression was not detected in smac KO mice.
The indicated MEFs or MEMs were exposed to 2 uM STS for 4 h and analyzed by Western blot. Accumulation of Smac/Diablo in mitochondrion-depleted cytosol fractions fromSTS-treated Apaf-1 KO cells were detected by anti-smac antibodies. Smac expression was not detected in smac KO mice.
Description Immunohistochemistry Validation of Smac in Human gastric carcinoma (Kim et al., 2011)
Smac was highly expressed in gastric mucosa of patients with gastric carcinoma.
Smac was highly expressed in gastric mucosa of patients with gastric carcinoma.
Description Immunofluorescence Analysis of Smac in NB4-LR1 Cells (Saumet et al., 2005)
NB4-LR1 cells were either treated with ATRA(1 uM) for 3 days without or with the T3C1 recombinant fragment (3 uM) or treated with staurosporine (STP; 5 uM ) for 3.5 hours. STP, but not ATRA or AYRA/T3C1 induced the release of smac.
NB4-LR1 cells were either treated with ATRA(1 uM) for 3 days without or with the T3C1 recombinant fragment (3 uM) or treated with staurosporine (STP; 5 uM ) for 3.5 hours. STP, but not ATRA or AYRA/T3C1 induced the release of smac.
Description Induced Expression Validation in Rat Liver (Genestier et al., 2005)
Mitochondria from rat liver were treated with increasing concentrations of rPVL (A), rLukS (B), or Bax alpha (C) for 1 hour at 30 C. rPVL induces the release of the apoptogenic proteins cytochrome c and Smac/DIABLO from isolated mitochondria.
Mitochondria from rat liver were treated with increasing concentrations of rPVL (A), rLukS (B), or Bax alpha (C) for 1 hour at 30 C. rPVL induces the release of the apoptogenic proteins cytochrome c and Smac/DIABLO from isolated mitochondria.
Description Overxpression Validation in HEK293T Cells (Flygare et al., 2012)
HEK293T cells were transiently transfected with Smac and Myc-tagged cIAP1, cIAP2, ML-IAP, or empty vector. Cells were lysed, and lysates were incubated with the indicated concentrations of 1 and immunoprecipitated with anti-Myc antibody (left panels). Samples were then immunoblotted with anti-Smac and anti-Myc antibodies. Whole-cell lysates are shown in the right panel.
HEK293T cells were transiently transfected with Smac and Myc-tagged cIAP1, cIAP2, ML-IAP, or empty vector. Cells were lysed, and lysates were incubated with the indicated concentrations of 1 and immunoprecipitated with anti-Myc antibody (left panels). Samples were then immunoblotted with anti-Smac and anti-Myc antibodies. Whole-cell lysates are shown in the right panel.
Handling
Storage This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Dilution Instructions Dilute in PBS or medium which is identical to that used in the assay system.
Application Instructions Immunoblotting: use at 1ug/mL.
Positive control: Mouse heart tissue lysate.
Immunohistochemistry: use at 2ug/mL.
These are recommended concentrations.
Enduser should determine optimal concentrations for their applications.
Positive control: Mouse heart tissue lysate.
Immunohistochemistry: use at 2ug/mL.
These are recommended concentrations.
Enduser should determine optimal concentrations for their applications.
References & Data Sheet
Data Sheet 
Download PDF Data Sheet
Download PDF Data Sheet