Name: Anti-Smac/Diablo antibody [Y12]
SKU: ab32023
Rating: 4 (5 reviews)

Rabbit Recombinant Monoclonal Smac/Diablo antibody. Suitable for IHC-P, IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 23 publications.

Images

Western blot - Anti-Smac/Diablo antibody [Y12] (AB32023), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Expected	Tested	Tested
Rat	Not recommended	Expected	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/6000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/80	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -
Species Mouse	Dilution info 1/100	Notes -
Species Rat	Dilution info 1/100	Notes -

Associated Products

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Target data

See full target information DIABLO

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 (PubMed:15200957, PubMed:36758104, PubMed:36758105, PubMed:36758106). Isoform 3. Attenuates the stability and apoptosis-inhibiting activity of XIAP/BIRC4 by promoting XIAP/BIRC4 ubiquitination and degradation through the ubiquitin-proteasome pathway. Also disrupts XIAP/BIRC4 interacting with processed caspase-9 and promotes caspase-3 activation. Isoform 1. Defective in the capacity to down-regulate the XIAP/BIRC4 abundance.

Alternative names

SMAC, DIABLO, Diablo IAP-binding mitochondrial protein, Direct IAP-binding protein with low pI, Second mitochondria-derived activator of caspases

Recommended products

Rabbit Recombinant Monoclonal Smac/Diablo antibody. Suitable for IHC-P, IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 23 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: Y12
Purification technique: Affinity purification Protein A
Epitope: ab32023 reacts with an epitope located in the C terminal region of Smac / Diablo.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Smac/Diablo is a mitochondrial protein that promotes apoptosis by interfering with inhibitor of apoptosis proteins (IAPs). The protein also known as Second Mitochondria-derived Activator of Caspases weighs approximately 27 kDa. Researchers detect Smac/Diablo in various tissues with higher expression levels in organs like the heart and brain. This protein's expression also varies depending on the cell's state and external stimuli.

Biological function summary

Smac/Diablo plays a significant role in programmed cell death by binding to IAPs and negating their inhibition of caspases the key executors of apoptosis. Smac/Diablo releases from mitochondria into the cytosol when apoptotic signals activate. It does not form complexes but interacts with IAPs facilitating the activation of caspases and enhancing the apoptotic response. Its interaction with IAPs highlights its important function in apoptosis regulation.

Pathways

Smac/Diablo integrates into the mitochondrial apoptosis pathway contributing to the intrinsic pathway of apoptosis. It closely interacts with proteins such as caspases and IAPs. This integration is important for apoptosis regulation linking mitochondrial outer membrane permeabilization with the activation of caspases. Additionally the protein engages in the cytochrome c pathway promoting apoptosis by restricting IAPs and aiding in the release of cytochrome c important for apoptosis progression.

Associated diseases and disorders

Researchers associate Smac/Diablo with cancer and neurodegenerative diseases like Alzheimer's disease. In cancer elevated Smac/Diablo levels can result in increased apoptotic death of cancerous cells potentially serving as a target for therapy. It also interacts with proteins like XIAP in these disease contexts. In neurodegenerative disorders dysregulation of apoptosis pathways involving Smac/Diablo might contribute to excessive neuronal cell death. Understanding Smac/Diablo's role in these diseases provides insights into possible therapeutic interventions.

Product promise

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11 product images

Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023)
All lanes: Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023) at 1/1000 dilution
All lanes: Jurkat cell lysate
Predicted band size: 27 kDa
Observed band size: 21 kDa
Immunoprecipitation - Anti-Smac/Diablo antibody [Y12] (ab32023)
Smac/Diablo was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 μg with ab32023 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10 μg
Lane 2: ab32023 IP in Jurkat whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32023 in Jurkat whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Smac/Diablo antibody [Y12] (ab32023)
Predicted band size: 27 kDa
Observed band size: 21 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Smac/Diablo antibody [Y12] (ab32023)
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Smac / Diablo with ab32023 at 1/500. Cells were fixed with 100% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Control: PBS only.
Nuclear counter stain: DAPI.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smac/Diablo antibody [Y12] (ab32023)
ab32023, at a 1/10 dilution, staining human Smac/Diablo in skin cancer tissue by immunohistochemistry, paraffin-embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Image from Lamers F et al. BMC Cancer. 2012 Jul 12;12:285. doi: 10.1186/1471-2407-12-285. Fig 3.; 12 July 2012 BMC Cancer 2012 12:285.
Immunocytochemistry/ Immunofluorescence - Anti-Smac/Diablo antibody [Y12] (ab32023)
Immunofluorescence analysis of IMR32 cells, staining Smac / Diablo with ab32023. Cells were fixed with 4% paraformaldehyde before incubating with primary antibody. An Alexa Fluor^® conjugated anti-rabbit IgG was used as the secondary antibody.
Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023)
Loading control: Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) (1/200000) (36KDa).

Blocking/Diluting buffer and concentration 5% NFDM/TBST
Lanes 1 - 4: Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-Smac/Diablo antibody [Y12] - BSA and Azide free (Anti-Smac/Diablo antibody [Y12] - BSA and Azide free ab239797)
Lane 1: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 3: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Lane 4: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 37s
Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023)
Loading control: Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) (1/200000) (36KDa).

Blocking/Diluting buffer and concentration 5% NFDM/TBST

Low expression: spleen, brain, heart (PMID: 12058276)
Lanes 1 - 8: Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023) at 1/1000 dilution
Lanes 1 - 8: Western blot - Anti-Smac/Diablo antibody [Y12] - BSA and Azide free (Anti-Smac/Diablo antibody [Y12] - BSA and Azide free ab239797)
Lane 1: Mouse testis tissue lysate at 20 µg
Lane 2: Mouse brain tissue lysate at 20 µg
Lane 3: Mouse heart tissue lysate at 20 µg
Lane 4: Mouse spleen tissue lysate at 20 µg
Lane 5: Rat testis tissue lysate at 20 µg
Lane 6: Rat brain tissue lysate at 20 µg
Lane 7: Rat heart tissue lysate at 20 µg
Lane 8: Rat spleen tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 10s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smac/Diablo antibody [Y12] (ab32023)
Immunohistochemistry analysis of paraffin-embedded mouse testis tissue sections labeling Smac/Diablo with ab32023 at 1/6000 dilution, incubated for 30 mins at room temperature followed by Rabbit specific IHC polymer detection kit HRP/DAB ab209101 (ready to use). Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smac/Diablo antibody [Y12] (ab32023)
Immunohistochemistry analysis of paraffin-embedded mouse brain tissue sections labeling Smac/Diablo with ab32023 at 1/6000 dilution, incubated for 30 mins at room temperature followed by Rabbit specific IHC polymer detection kit HRP/DAB ab209101 (ready to use). Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Positive staining on mouse brain.
Immunocytochemistry/ Immunofluorescence - Anti-Smac/Diablo antibody [Y12] (ab32023)
Immunocytochemistry/Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labelling Smac / Diablo with ab32023 at 1/100 dilution (green). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (1/1000) was used as the secondary antibody. Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker at 1/1000 was as the counterstain antibody followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (magenta). Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Negative Control 1: ab32023 at 1/100 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000.
Negative Control 2: anti-COX IV antibody at 1/1000 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at 1/1000.

Confocal image showing positive staining in mitochondria of C6 cell line (shown in green).
Immunocytochemistry/ Immunofluorescence - Anti-Smac/Diablo antibody [Y12] (ab32023)
Immunocytochemistry/Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labelling Smac / Diablo with ab32023 at 1/100 dilution (green). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (1/1000) was used as the secondary antibody. Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker at 1/1000 was as the counterstain antibody followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (magenta). Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Negative Control 1: ab32023 at 1/100 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/1000.
Negative Control 2: anti-COX IV antibody at 1/1000 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at 1/1000.
Confocal image showing positive staining in mitochondria of NIH/3T3 cell line (shown in green).