Recombinant Anti-Smac/Diablo antibody [Y12] (ab32023)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y12] to Smac/Diablo
- Suitable for: WB, IHC-P, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Smac/Diablo antibody [Y12]
See all Smac/Diablo primary antibodies -
Description
Rabbit monoclonal [Y12] to Smac/Diablo -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide within Human Smac/ Diablo (C terminal). The exact sequence is proprietary.
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Epitope
ab32023 reacts with an epitope located in the C terminal region of Smac / Diablo. -
Positive control
- WB: Jurkat whole cell lysate (ab7899). IHC-P: Human skin cancer tissue. ICC/IF: HeLa cells. IP: Jurkat.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y12 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32023 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (3) |
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 27 kDa).
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IHC-P |
1/10. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP | (1) |
1/10 - 1/80.
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ICC/IF |
1/500.
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Notes |
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WB
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 27 kDa). |
IHC-P
1/10. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
1/10 - 1/80. |
ICC/IF
1/500. |
Target
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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). -
Tissue specificity
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. -
Domain
The mature N-terminus mediates interaction with XIAP. -
Cellular localization
Mitochondrion. Released into the cytosol when cells undergo apoptosis. - Information by UniProt
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Database links
- Entrez Gene: 56616 Human
- Entrez Gene: 66593 Mouse
- Entrez Gene: 288753 Rat
- Omim: 605219 Human
- SwissProt: Q9NR28 Human
- SwissProt: Q9JIQ3 Mouse
- Unigene: 169611 Human
- Unigene: 46716 Mouse
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Alternative names
- 0610041G12Rik antibody
- DBLOH_HUMAN antibody
- DBOH antibody
see all
Images
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Anti-Smac/Diablo antibody [Y12] (ab32023) at 1/1000 dilution + Jurkat cell lysate
Predicted band size: 27 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Smac / Diablo with ab32023 at 1/500. Cells were fixed with 100% methanol. 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.
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Immunocytochemistry/ Immunofluorescence - Anti-Smac/Diablo antibody [Y12] (ab32023)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
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.
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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) (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 (ab172730) instead of ab32023 in Jurkat whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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Immunoprecipitation - Anti-Smac/Diablo antibody [Y12] (ab32023)This image is courtesy of an anonymous Abreview.
Smac / Diablo immunprecipitated from mitochondria isolated from mice brains.
Total protein in input: 800 µg.
Immunoprecipitation step: Dynabeads protein G.
Incubation: 16 hours, 4°C.
Diluent: TNE buffer.For western blot, ab32023 was used at a 1/1000 dilution.
Lanes:
1. BioRad Precision Plus Protein standards, bands from top to bottom: 250, 150, 100, 75, 50, 37, 25, 20, 15, 10 kDa
2. Mitochondria isolated from mice brains, lysate
3. IP with anti-SMAC/DIABLO ab32023
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (20)
ab32023 has been referenced in 20 publications.
- McKenna S et al. BAX and SMAC regulate bistable properties of the apoptotic caspase system. Sci Rep 11:3272 (2021). PubMed: 33558564
- Zhou H et al. Upconversion NIR-II fluorophores for mitochondria-targeted cancer imaging and photothermal therapy. Nat Commun 11:6183 (2020). PubMed: 33273452
- Xian H & Liou YC Loss of MIEF1/MiD51 confers susceptibility to BAX-mediated cell death and PINK1-PRKN-dependent mitophagy. Autophagy 15:2107-2125 (2019). PubMed: 30894073
- Wu B et al. Combination of Sodium Selenite and Doxorubicin Prodrug Ac-Phe-Lys-PABC-ADM Affects Gastric Cancer Cell Apoptosis in Xenografted Mice. Biomed Res Int 2019:2486783 (2019). PubMed: 31531348
- Wei Q et al. The mitochondrial pathway is involved in sodium fluoride (NaF)-induced renal apoptosis in mice. Toxicol Res (Camb) 7:792-808 (2018). PubMed: 30310657