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

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Western blot - 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

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

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

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y12
Isotype

IgG
Research areas

Alternative Versions
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- Jurkat whole cell lysate (ab7899)
Recombinant Protein
- Recombinant Human Smac/Diablo protein (His tag) (ab219716)
- Recombinant Human Smac/Diablo protein (ab87721)

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
WB	(3)	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)	1/10 - 1/80.
ICC/IF		1/500.

Notes
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.

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.
Target information above from: UniProt accession Q9NR28 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
Alternative names
- 0610041G12Rik antibody
- DBLOH_HUMAN antibody
- DBOH antibody
- DFNA64 antibody
- diablo antibody
- Diablo homolog (Drosophila) antibody
- Diablo homolog antibody
- Diablo homolog mitochondrial antibody
- Diablo IAP binding mitochondrial protein antibody
- Diablo like protein antibody
- DIABLO S antibody
- Direct IAP binding protein with low pI antibody
- Direct IAP-binding protein with low pI antibody
- FLJ10537 antibody
- FLJ25049 antibody
- mitochondrial antibody
- Mitochondrial Smac protein antibody
- Second mitochondria derived activator of caspase antibody
- Second mitochondria-derived activator of caspase antibody
- second mitochondrial activator of caspases antibody
- SMAC 3 antibody
- Smac antibody
- Smac protein antibody
- SMAC3 antibody
see all

Western blot - Anti-Smac/Diablo antibody [Y12] (ab32023)

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 - 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. 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.
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.
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) (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.
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
See Abreview
Anti-Smac/Diablo antibody [Y12] (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.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab32023? Please let us know so that we can cite the reference in this datasheet.

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

View all Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Application

Immunoprecipitation

Sample

Mouse Tissue lysate - other (Mitochondria isolated from mice brains, lysed by t)

Total protein in input

800 µg

Immuno-precipitation step

Other - Dynabeads protein G

Specification

Mitochondria isolated from mice brains, lysed by t

Abcam user community

Verified customer

Submitted Mar 05 2019

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Reduced Denaturing (15% Gel)

Sample

Mouse Tissue lysate - other (Skeletal muscle cytosolic fractions)

Specification

Skeletal muscle cytosolic fractions

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 25 2015

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Reduced Denaturing (15% Gel)

Sample

Rat Tissue lysate - other (Skeletal muscle cytosolic fractions)

Specification

Skeletal muscle cytosolic fractions

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 19 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (Platelets)

Loading amount

20 µg

Specification

Platelets

Treatment

ADP for 30 min

Gel Running Conditions

Reduced Denaturing (4-20%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 23 2011

Application

Flow Cytometry

Sample

Human Cell (Platelets)

Specification

Platelets

Preparation

Cell harvesting/tissue preparation method: PL were isolated spinning Platelet rich plasma on Histopaque
Sample buffer: PBS

Fixation

Paraformaldehyde

Permeabilization

Yes - 0.1% Triton-X100 in 2% BSA for 15min

Gating Strategy

Platelets

Abcam user community

Verified customer

Submitted Dec 31 2010

Question

Dear sirs

Please I need a secondary antibody working with ab32023 on flowcytometer by dye (FITC and PE) please suggest

Sincerely yours,

Abcam community

Verified customer

Asked on Mar 05 2012

Answer

Thank you for your inquiry.

I chose two secondary antibodies for each conjugate. If you would like to review the whole selection of possible secondary antibodies,please use our secondary antibody search tool:

https://www.abcam.com/index.html?pageconfig=productmap&cl=918

anti rabbit conjugated with FITC:

ab98502
https://www.abcam.com/Donkey-Rabbit-IgG-HL-FITC-preadsorbed-ab98502.html (or use the following: https://www.abcam.com/Donkey-Rabbit-IgG-HL-FITC-preadsorbed-ab98502.html).

ab98466
https://www.abcam.com/Goat-Rabbit-IgG-Fc-FITC-preadsorbed-ab98466.html (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-Fc-FITC-preadsorbed-ab98466.html).

anti rabbit conjugated with PE:

ab97070
https://www.abcam.com/Goat-Rabbit-IgG-HL-PE-preadsorbed-ab97070.html (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-HL-PE-preadsorbed-ab97070.html).

ab98494
https://www.abcam.com/Donkey-Fab2-Rabbit-IgG-HL-PE-pre-adsorbed-ab98494.html (or use the following: https://www.abcam.com/Donkey-Fab2-Rabbit-IgG-HL-PE-pre-adsorbed-ab98494.html).

I can recommend to also review the new conjugates which have are much more stable than the old conjugates FITC and PE:

https://www.abcam.com/index.html?pageconfig=resource&rid=12934

For more general information please follow the following links:

https://www.abcam.com/index.html?pageconfig=resource&rid=12922

https://www.abcam.com/index.html?pageconfig=resource&rid=12825

https://www.abcam.com/index.html?pageconfig=resource&rid=12722

https://www.abcam.com/index.html?pageconfig=resource&rid=12956

I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

Abcam Scientific Support

Answered on Mar 05 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Epitope

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (20)

Customer reviews and Q&As

Immunoprecipitation abreview for Anti-Smac / Diablo antibody [Y12]

Western blot abreview for Anti-Smac / Diablo antibody [Y12]

Western blot abreview for Anti-Smac / Diablo antibody [Y12]

Western blot abreview for Anti-Smac / Diablo antibody [Y12]

Flow Cytometry abreview for Anti-Smac / Diablo antibody [Y12]