Overview

  • Product name
  • Description
    Rabbit polyclonal to APAF1
  • Tested applications
    Suitable 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 notes
    Apoptosis Protease Activating Factor.

Properties

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

  • Function
    Oligomeric 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 specificity
    Ubiquitous. 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 similarities
    Contains 1 CARD domain.
    Contains 1 NB-ARC domain.
    Contains 13 WD repeats.
  • Domain
    The CARD domain mediates interaction with APIP.
  • Cellular localization
    Cytoplasm.
  • 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:
  • Somarajan BI  et al. Rapid Eye Movement Sleep Deprivation Induces Neuronal Apoptosis by Noradrenaline Acting on Alpha1 Adrenoceptor and by Triggering Mitochondrial Intrinsic Pathway. Front Neurol 7:25 (2016). Read more (PubMed: 27014180) »
  • 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) »

See all 4 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

Submitted Feb 09 2016

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
Username

Abcam user community

Verified customer

Submitted Jun 29 2015

Thank you for your reply.

As requested, I have placed a free of charge vial of ab59388 onto order 1083546. This should be shipped today or tomorrow and estimated delivery time is next Wednesday or Thursday.

I hope this will be hel...

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According to our records, ab59388 was ordered in October which is past our 6 month guarantee period. However, reviewing the details, I can make an exception and provide a re...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be plea...

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Thank you for providing this further information.

I appreciate your cooperation and understand your concerns. It is regrettable the results have not been succesful. I have traced your previous corresopndence and reveiwed this case.
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Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this...

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Thank you again for your enquiry. Regarding ab4969, ab4957, and ab4958 (the peptides corresponding to the antibodies you indicated), they are all greater than 95% pure, and they were purified via HPLC. If you have any further questions, please conta...

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Thank you for your enquiry. For ab2001 and ab7346, the peptides are non-HPLC, which averages out to 60-70% purity. Ab2001 (the antibody) was immunoaffinity chromatography purified IgG, and ab7346 (CCR5 antibody) is purified IgG. I will have to get b...

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