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: Mouse, Rat, Human
Product nameAnti-Smac/Diablo antibody [Y12]
See all Smac/Diablo primary antibodies
DescriptionRabbit monoclonal [Y12] to Smac/Diablo
Tested applicationsSuitable for: WB, IHC-P, IP, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Smac/ Diablo (C terminal). The exact sequence is proprietary.
Epitopeab32023 reacts with an epitope located in the C terminal region of Smac / Diablo.
- WB: Jurkat whole cell lysate (ab7899). IHC-P: Human skin cancer tissue. ICC/IF: HeLa cells.
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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.|
FunctionPromotes 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 specificityUbiquitously 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.
DomainThe mature N-terminus mediates interaction with XIAP.
Cellular localizationMitochondrion. Released into the cytosol when cells undergo apoptosis.
- Information by UniProt
- 0610041G12Rik antibody
- DBLOH_HUMAN antibody
- DBOH antibody
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.
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.
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.
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.
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
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.
ab32023 has been referenced in 16 publications.
- Jiang Y et al. MAP30 promotes apoptosis of U251 and U87 cells by suppressing the LGR5 and Wnt/ß-catenin signaling pathway, and enhancing Smac expression. Oncol Lett 15:5833-5840 (2018). PubMed: 29556310
- Zhang W et al. Long non-coding RNA-ROR aggravates myocardial ischemia/reperfusion injury. Braz J Med Biol Res 51:e6555 (2018). PubMed: 29694511
- 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
- Liu R et al. A New Perspective for Osteosarcoma Therapy: Proteasome Inhibition by MLN9708/2238 Successfully Induces Apoptosis and Cell Cycle Arrest and Attenuates the Invasion Ability of Osteosarcoma Cells in Vitro. Cell Physiol Biochem 41:451-465 (2017). PubMed: 28214890
- Giampazolias E et al. Mitochondrial permeabilization engages NF-?B-dependent anti-tumour activity under caspase deficiency. Nat Cell Biol 19:1116-1129 (2017). WB . PubMed: 28846096