Recombinant
RabMAb

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

Reviews (5)Q&A (1)References (13)

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, Flow Cyt, 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

    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 rabbit monoclonal antibody.

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 Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP 1/10 - 1/80.
Flow Cyt 1/10 - 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.

References

This product has been referenced in:

  • 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). Read more (PubMed: 29556310) »
  • Zhang W  et al. Long non-coding RNA-ROR aggravates myocardial ischemia/reperfusion injury. Braz J Med Biol Res 51:e6555 (2018). Read more (PubMed: 29694511) »
See all 13 Publications for this product

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

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

Sign up