Anti-APAF1 Rabbit Monoclonal Antibody for WB and ELISA - O14727

Core Product Specifications and Parameters

Biological Background: APAF1 Function and Localization

• APAF1 (Apoptotic protease-activating factor 1), also known as CED4 or APAF-1, is encoded by the APAF1 gene (KIAA0413) and exists as multiple isoforms due to alternative splicing.
• The protein functions as a central component of the apoptosome, where oligomeric APAF1 mediates cytochrome c-dependent autoproteolytic activation of pro-caspase-9, subsequently activating caspase-3 to execute apoptosis; this process requires ATP binding.
• APAF1 is a multi-domain protein containing an N-terminal caspase recruitment domain (CARD), a central nucleotide-binding and oligomerization domain (NOD), and C-terminal WD40 repeat regions critical for cytochrome c binding.
• Subcellularly, APAF1 resides predominantly in the cytoplasm under normal conditions, and upon apoptosis induction, it translocates to the apoptosome complex.
• Expression is ubiquitous, with highest levels detected in adult spleen, peripheral blood leukocytes, fetal brain, kidney, and lung; isoform 1 is specifically found in heart, kidney, and liver.
• The protein is subject to ATP/dATP binding and likely calcium-mediated regulation, as indicated by keyword associations, and features WD repeats essential for protein-protein interactions.
• Keywords: 3D-structure, Alternative splicing, Apoptosis, ATP-binding, Calcium, Cytoplasm, Direct protein sequencing, Nucleotide-binding, Proteomics identification, Reference proteome, Repeat, WD repeat.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 470-600 of human APAF1 (central NOD region), ensuring detection of full-length and truncation variants, but specificity should be verified in relevant cell models.
• For Western blotting, consider that APAF1 migrates at multiple molecular weights (38-142 kDa) due to isoform diversity and caspase cleavage; band pattern may vary with cell type and apoptotic stimuli.
• ELISA applications can be developed using a recombinant APAF1 standard curve; include appropriate positive (e.g., staurosporine-treated Jurkat lysates) and negative controls to validate assay specificity.
• Cross-reactivity with mouse APAF1 is expected based on sequence homology, but empirical confirmation in mouse tissues or cell lines is recommended.
• Optimization of antibody dilution, incubation conditions, and detection reagents is advised for each experimental system.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your integrated sourcing partner for IVD raw materials, providing validated antibodies, antigen pairs, and ancillary reagents from discovery to commercial scale. This anti-APAF1 monoclonal antibody supports apoptosis research and diagnostic assay development, with reliable bulk supply and technical consultation available. For rare targets or customized solutions, CamelBio leverages global networks to bridge supply gaps efficiently.