Recombinant
RabMAb

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

Reviews (5)Q&A (1)References (16)

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

Overview

  • 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: Mouse, Rat, Human

  • 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.

  • 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.

Properties

Applications

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 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

  • 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

  • Database links

    see all

  • 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

Images

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

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.

Click here to view the general protocols

References (16)

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

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
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-6 of 6 Abreviews or Q&A

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

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Mar 05 2019

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

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Feb 25 2015

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

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Feb 19 2015

Question

Dear sirs

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



Sincerely yours,

Read More
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/index.html?datasheet=98502 (or use the following: https://www.abcam.com/index.html?datasheet=98502).

ab98466
https://www.abcam.com/index.html?datasheet=98466 (or use the following: https://www.abcam.com/index.html?datasheet=98466).

anti rabbit conjugated with PE:

ab97070
https://www.abcam.com/index.html?datasheet=97070 (or use the following: https://www.abcam.com/index.html?datasheet=97070).

ab98494
https://www.abcam.com/index.html?datasheet=98494 (or use the following: https://www.abcam.com/index.html?datasheet=98494).

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.

Read More

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

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Nov 23 2011

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

 Average
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Dec 31 2010

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com