Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human APAF1 (N terminal).
WB: HeLa cell lysate.
IHC-P: Stomach adenocarcinoma.
FC: HeLa cells
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 135 kDa (predicted molecular weight: 141 kDa).
For unpurified, use 1/50 - 1/100.
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.
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.
Western blot - Anti-APAF1 antibody [E38] (ab32372)
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
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.
Western blot - Anti-APAF1 antibody [E38] (ab32372)
Anti-APAF1 antibody [E38] (ab32372) at 1/500 dilution (unpurified) + HeLa cell lysate
Unpurified ab32372, at a dilution of 1/50, staining APAF1 in paraffin embedded stomach adenocarcinoma by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Anti-APAF1 antibody [E38] (ab32372)Image courtesy of an anonymous Abreview.
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.
References for Anti-APAF1 antibody [E38] (ab32372)
This product has been referenced in:
Wang C et al. Bifidobacterial recombinant thymidine kinase-ganciclovir gene therapy system induces FasL and TNFR2 mediated antitumor apoptosis in solid tumors. BMC Cancer16:545 (2016).
Read more (PubMed: 27464624) »
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 One10:e0120217 (2015).
Read more (PubMed: 25781629) »