Product: APAF1 Antibody
Catalog: AF0117
Description: Rabbit polyclonal antibody to APAF1
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 30~40kDa(ALT), 120~140kDa; 142kD(Calculated).
Uniprot: O14727
RRID: AB_2833267

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:3000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
APAF1 Antibody detects endogenous levels of total APAF1.
RRID:
AB_2833267
Cite Format: Affinity Biosciences Cat# AF0117, RRID:AB_2833267.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

APAF 1; Apaf-1; APAF_HUMAN; Apaf1; Apoptotic peptidase activating factor 1; Apoptotic protease activating factor 1; Apoptotic protease activating factor; Apoptotic protease-activating factor 1; CED 4; CED4; KIAA0413;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O14727 APAF_HUMAN:

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.

Description:
APAF 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. Induced by E2F and p53 in apoptotic neurons. Monomer. Oligomerizes upon binding of cytochrome c and dATP. Oligomeric Apaf-1 and pro-caspase-9 bind to each other via their respective NH2-terminal CARD domains and consecutively mature caspase-9 is released from the complex. Pro-caspase-3 is recruited into the Apaf-1-pro-caspase-9 complex via interaction with pro-caspase-9. Interacts with APIP. 6 isoforms of the human protein are produced by alternative splicing.
Sequence:
MDAKARNCLLQHREALEKDIKTSYIMDHMISDGFLTISEEEKVRNEPTQQQRAAMLIKMILKKDNDSYVSFYNALLHEGYKDLAALLHDGIPVVSSSSGKDSVSGITSYVRTVLCEGGVPQRPVVFVTRKKLVNAIQQKLSKLKGEPGWVTIHGMAGCGKSVLAAEAVRDHSLLEGCFPGGVHWVSVGKQDKSGLLMKLQNLCTRLDQDESFSQRLPLNIEEAKDRLRILMLRKHPRSLLILDDVWDSWVLKAFDSQCQILLTTRDKSVTDSVMGPKYVVPVESSLGKEKGLEILSLFVNMKKADLPEQAHSIIKECKGSPLVVSLIGALLRDFPNRWEYYLKQLQNKQFKRIRKSSSYDYEALDEAMSISVEMLREDIKDYYTDLSILQKDVKVPTKVLCILWDMETEEVEDILQEFVNKSLLFCDRNGKSFRYYLHDLQVDFLTEKNCSQLQDLHKKIITQFQRYHQPHTLSPDQEDCMYWYNFLAYHMASAKMHKELCALMFSLDWIKAKTELVGPAHLIHEFVEYRHILDEKDCAVSENFQEFLSLNGHLLGRQPFPNIVQLGLCEPETSEVYQQAKLQAKQEVDNGMLYLEWINKKNITNLSRLVVRPHTDAVYHACFSEDGQRIASCGADKTLQVFKAETGEKLLEIKAHEDEVLCCAFSTDDRFIATCSVDKKVKIWNSMTGELVHTYDEHSEQVNCCHFTNSSHHLLLATGSSDCFLKLWDLNQKECRNTMFGHTNSVNHCRFSPDDKLLASCSADGTLKLWDATSANERKSINVKQFFLNLEDPQEDMEVIVKCCSWSADGARIMVAAKNKIFLFDIHTSGLLGEIHTGHHSTIQYCDFSPQNHLAVVALSQYCVELWNTDSRSKVADCRGHLSWVHGVMFSPDGSSFLTSSDDQTIRLWETKKVCKNSAVMLKQEVDVVFQENEVMVLAVDHIRRLQLINGRTGQIDYLTEAQVSCCCLSPHLQYIAFGDENGAIEILELVNNRIFQSRFQHKKTVWHIQFTADEKTLISSSDDAEIQVWNWQLDKCIFLRGHQETVKDFRLLKNSRLLSWSFDGTVKVWNIITGNKEKDFVCHQGTVLSCDISHDATKFSSTSADKTAKIWSFDLLLPLHELRGHNGCVRCSAFSVDSTLLATGDDNGEIRIWNVSNGELLHLCAPLSEEGAATHGGWVTDLCFSPDGKMLISAGGYIKWWNVVTGESSQTFYTNGTNLKKIHVSPDFKTYVTVDNLGILYILQTLE

PTMs - O14727 As Substrate

Site PTM Type Enzyme
Y24 Phosphorylation
S31 Phosphorylation
T48 Phosphorylation
K63 Ubiquitination
K131 Ubiquitination
K139 Ubiquitination
K144 Ubiquitination
S161 Phosphorylation
K189 Ubiquitination
K192 Ubiquitination
K198 Ubiquitination
T204 Phosphorylation
K224 Ubiquitination
S238 Phosphorylation
S248 Phosphorylation
K267 Ubiquitination
S268 Phosphorylation Q15418 (RPS6KA1)
S272 Phosphorylation
K277 Ubiquitination
Y278 Phosphorylation
K288 Ubiquitination
K303 Ubiquitination
S320 Phosphorylation
K348 Ubiquitination
S357 Phosphorylation Q15418 (RPS6KA1)
S369 Phosphorylation
S371 Phosphorylation
K380 Ubiquitination
Y382 Phosphorylation
Y383 Phosphorylation
K391 Ubiquitination
K458 Ubiquitination
S506 Phosphorylation
K581 Ubiquitination
K600 Ubiquitination
K601 Ubiquitination
K637 Ubiquitination
K643 Acetylation
K643 Ubiquitination
K649 Ubiquitination
K654 Acetylation
K733 Ubiquitination
K756 Ubiquitination
K768 Ubiquitination
K818 Ubiquitination
S901 Phosphorylation
T905 Phosphorylation
K916 Ubiquitination
K1048 Ubiquitination
K1077 Ubiquitination
K1079 Ubiquitination
K1099 Ubiquitination
T1103 Phosphorylation
K1107 Ubiquitination
T1108 Phosphorylation
K1222 Ubiquitination

Research Backgrounds

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.

Subcellular Location:

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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.

Subunit Structure:

Monomer. Oligomerizes to a heptameric ring, known as the apoptosome, upon binding of cytochrome c and dATP. Oligomeric Apaf-1 and pro-caspase-9 bind to each other via their respective NH2-terminal CARD domains and consecutively mature caspase-9 is released from the complex. Pro-caspase-3 is recruited into the Apaf-1-pro-caspase-9 complex via interaction with pro-caspase-9. Interacts with APIP. Interacts (via CARD and NACHT domains) with NAIP/BIRC1 (via NACHT domain). Interacts with CIAO2A.

Family&Domains:

The CARD domain mediates interaction with APIP.

The monomeric form is autoinhibited in a closed conformation through a bound ADP at the nucleotide binding site. Exchange of ADP for ATP and binding of cytochrome c trigger a large conformational change where the first WD repeat region swings out, allowing the NB-ARC domain to rotate and expose the contact areas for oligomerization (By similarity).

Research Fields

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis - multiple species.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Human Diseases > Neurodegenerative diseases > Parkinson's disease.

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Neurodegenerative diseases > Huntington's disease.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer.   (View pathway)

References

1). Ai X et al. Berberis dictyophylla F. inhibits angiogenesis and apoptosis of diabetic retinopathy via suppressing HIF-1α/VEGF/DLL-4/Notch-1 pathway. Journal of Ethnopharmacology 2022 Oct 5;296:115453. (PubMed: 35697191) [IF=5.4]

2). Ding Y et al. Carnitine palmitoyltransferase 1 (CPT1) alleviates oxidative stress and apoptosis of hippocampal neuron in response to beta-Amyloid peptide fragment Aβ25-35. Bioengineered 2021 Dec;12(1):5440-5449. (PubMed: 34424821) [IF=4.9]

Application: WB    Species: Mice    Sample: Aβ25-35-induced HT22 cells

Figure 4. CPT1C overexpression decreased the apoptosis of Aβ25-35-induced HT22 cells. Following transfection of Ov-CPT1C or Ov-NC for 24 h, HT22 cells were treated with Aβ25–35 for another 24 h (a) The apoptosis of Aβ25-35-induced HT22 cells was evaluated by TUNEL. (b) The expressions of Bcl2, Bax, cleaved PARP and APAF-1 were measured by western blot. ***P < 0.001 vs. Control group, ###P < 0.001 vs A25-35 + Ov-NC.

3). Zhang SH et al. Ferulic acid ameliorates pentylenetetrazol-induced seizures by reducing neuron cell death. EPILEPSY RESEARCH 2019 Aug 5;156:106183 (PubMed: 31404716) [IF=2.2]

Application: WB    Species: rat    Sample: hippocampus

Fig. 5. |Western blot results showing the effect of FA on Apaf-1, caspase-9, caspase-3, Bcl-2, Bid, Bax, cleaved caspase-3 and cytochrome C expression in the hippocampus of rats with seizures. Western blot results show the expression of all of the measured proteins (A).

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