Overview

  • Product nameAnti-APAF1 antibody
    See all APAF1 primary antibodies
  • Description
    Rabbit polyclonal to APAF1
  • Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human APAF1 aa 12-28 (N terminal).
    Sequence:

    HREALEKDIKTSYIMDHC


    (Peptide available as ab7873)

  • Positive control
    • HeLa whole cell lysate. This antibody gave a positive result in IF/ICC when used in the following formaldehyde fixed cell lines: HeLa.
  • General notesApoptosis Protease Activating Factor.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab2001 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
IHC-P Use a concentration of 1 µg/ml.
ICC/IF Use a concentration of 10 µg/ml.
WB 1/1000 - 1/2000. Detects a band of approximately 141 kDa.Can be blocked with APAF1 peptide (ab7873).

Target

  • FunctionOligomeric Apaf-1 mediates the cytochrome c-dependent autocatalytic activation of pro-caspase-9 (Apaf-3), leading to the activation of caspase-3 and apoptosis. This activation requires ATP. Isoform 6 is less effective in inducing apoptosis.
  • Tissue specificityUbiquitous. Highest levels of expression in adult spleen and peripheral blood leukocytes, and in fetal brain, kidney and lung. Isoform 1 is expressed in heart, kidney and liver.
  • Sequence similaritiesContains 1 CARD domain.
    Contains 1 NB-ARC domain.
    Contains 13 WD repeats.
  • DomainThe CARD domain mediates interaction with APIP.
  • Cellular localizationCytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • APAF 1 antibody
    • Apaf-1 antibody
    • APAF_HUMAN antibody
    • Apaf1 antibody
    • Apoptotic peptidase activating factor 1 antibody
    • Apoptotic protease activating factor 1 antibody
    • Apoptotic protease activating factor antibody
    • Apoptotic protease-activating factor 1 antibody
    • CED 4 antibody
    • CED4 antibody
    • KIAA0413 antibody
    see all

Anti-APAF1 antibody images

  • All lanes : Anti-APAF1 antibody (ab2001) at 1 µg/ml

    Lane 1 : K562 cell lysate in absence of blocking peptide
    Lane 2 : K562 cell lysate in presence of blocking peptide.


    Observed band size : 141 kDa (why is the actual band size different from the predicted?)
  • ab2001 at 1µg/ml staining Apaf1 in human heart tissue by IHC 
  • ab2001 staining APAF1 in rat intestinal epithelial cells (RIE-1) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 1.5% serum for 60 minutes at 25°C. Samples were incubated with primary antibody (1/400 with 1x HBSS + 0.02% TritonX-100 + 1.5% FBS) for 3 hour at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/2000.

  • ICC/IF image of ab2001 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab2001 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab2001 staining APAF1 in mouse embryonic fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% milk for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 1X PBS + 0.02% Tween-20 + 10% milk) for 3 hours at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

References for Anti-APAF1 antibody (ab2001)

This product has been referenced in:
  • Cobbold LC  et al. Identification of internal ribosome entry segment (IRES)-trans-acting factors for the Myc family of IRESs. Mol Cell Biol 28:40-9 (2008). WB . Read more (PubMed: 17967896) »
  • Potokar M  et al. Subcellular localization of Apaf-1 in apoptotic rat pituitary cells. Am J Physiol Cell Physiol 290:C672-7 (2006). Read more (PubMed: 16207793) »

See all 3 Publications for this product

Product Wall

Application IHC - Wholemount
Sample Mouse Tissue (Transplanted lung tumor section, IHC-P (paraffin e)
Specification Transplanted lung tumor section, IHC-P (paraffin e
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Submitted Mar 11 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (rat intestinal epithelial cells (RIE-1))
Permeabilization Yes - 0.2% TritonX-100
Specification rat intestinal epithelial cells (RIE-1)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Feb 11 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Chinese Hamster Cell (Chinese hamster ovary cell)
Permeabilization Yes - 0.2% TritonX-100
Specification Chinese hamster ovary cell
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Feb 09 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Chinese Hamster Cell lysate - whole cell (Chinese hamster ovary cell)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 100 nM Bortezomib for 16 hours
Specification Chinese hamster ovary cell
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 09 2016

Application Western blot
Sample Rat Cell lysate - whole cell (rat intestinal epithelial cells (RIE-1))
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 100 nM Bortezomib for 16 hours
Specification rat intestinal epithelial cells (RIE-1)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 08 2016

Application Flow Cytometry
Sample Mouse Cell (Mouse Lewis Lung Carcinoma cells)
Permeabilization Yes - 0.2% Triton X-100
Gating Strategy No gating
Specification Mouse Lewis Lung Carcinoma cells
Preparation Cell harvesting/tissue preparation method: trypsin
Sample buffer: Hanks’ Balanced Salt solution (HBSS)
Fixation Paraformaldehyde
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Submitted Jan 21 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse embryonic fibroblasts)
Specification mouse embryonic fibroblasts
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Jul 22 2015

Application IHC - Wholemount
Sample Human Tissue (Human ovarian thecoma)
Specification Human ovarian thecoma
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Submitted Jun 30 2015

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS-7 cells)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 30 nM Bortezomib for 24 hours
Specification COS-7 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Jun 29 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (Lewis lung cancer, FL5.12, embryonic fibroblasts)
Gel Running Conditions Reduced Denaturing (4-20% Bis-Tris)
Loading amount 100000 cells
Specification Lewis lung cancer, FL5.12, embryonic fibroblasts
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted May 11 2015

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