Overview

  • Product nameAnti-APAF1 antibody [E38]
    See all APAF1 primary antibodies
  • Description
    Rabbit monoclonal [E38] to APAF1
  • Tested applicationsSuitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human APAF1 (N terminal).

  • Positive control
    • WB: HeLa cell lysate. IHC-P: Stomach adenocarcinoma. FC: HeLa cells
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab32372 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
ICC/IF 1/100.
Flow Cyt 1/30.
WB 1/1000. Detects a band of approximately 135 kDa (predicted molecular weight: 141 kDa).
IHC-P 1/250.

For unpurified, use 1/50 - 1/100.

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 [E38] images

  • All lanes : Anti-APAF1 antibody [E38] (ab32372) at 1/1000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : HT-1080 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 141 kDa
    Observed band size : 135 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human bladder cancer with purified ab32372 at a working dilution of 1/250. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Immunofluorescence staining of HeLa cells with purified ab32372 at a working dilution of 1 in 100, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab323722 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling APAF1 with purified ab32372 at 1/30 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Anti-APAF1 antibody [E38] (ab32372) at 1/1000 dilution (purified) + Mouse spleen tissue lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 141 kDa
    Observed band size : 135 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded mouse colon with purified ab32372 at a working dilution of 1/250. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Anti-APAF1 antibody [E38] (ab32372) at 1/500 dilution (unpurified) + HeLa cell lysate

    Predicted band size : 141 kDa
    Observed band size : 135 kDa (why is the actual band size different from the predicted?)
  • Unpurified ab32372, at a dilution of 1/50, staining APAF1 in paraffin embedded stomach adenocarcinoma by Immunohistochemistry.

  • Unpurified ab32372 staining APAF1 in human leukocytes by Immunocytochemistry/ Immunofluorescence.Cells were fixed in formaldehyde, permeabilized using 0.3% Triton X-100, blocked with 2% serum for 2 hours at 25°C, and then incubated with ab32372 at a 1/250 dilution for 9 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution.

    See Abreview

References for Anti-APAF1 antibody [E38] (ab32372)

This product has been referenced in:
  • Chen Q  et al. The transcription factor c-Myc suppresses MiR-23b and MiR-27b transcription during fetal distress and increases the sensitivity of neurons to hypoxia-induced apoptosis. PLoS One 10:e0120217 (2015). WB ; Mouse . Read more (PubMed: 25781629) »
  • Zhang W  et al. ELL inhibits E2F1 transcriptional activity by enhancing E2F1 deacetylation via recruitment of histone deacetylase 1. Mol Cell Biol 34:765-75 (2014). Read more (PubMed: 24344198) »

See all 8 Publications for this product

Product Wall

Application Western blot
Loading amount 100000 cells
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Jurkat)
Specification Jurkat
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Abcam user community

Verified customer

Submitted Feb 13 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (leukocytes)
Specification leukocytes
Fixative Formaldehyde
Permeabilization Yes - 0.3% Triton X-100
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 02 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (bone marrow)
Specification bone marrow
Fixative Methanol
Permeabilization No
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 19 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"