EnCor Biotechnology
Chicken Polyclonal Antibody to ACIII Cat# CPCA-ACIII
Description
The CPCA-ACIII antibody was made against the extreme C-terminal peptide of rat ACIII, PAAFPNGSSVTLPHQVVDNP, amino acids 1125-1144 of the Genbank entry NP_570135.2 coupled to MBS-activated keyhole limpet hemocyanin. The antibody works on mouse cells which express the same peptide and also on human cells, presumably because the corresponding peptide in the human AC3 sequence is the closely related peptide LATFPNGPSVTLPHQVVDNS. The antibody works well to identify neuronal cilia on human and rodent cells in IF and ICC, but is not recommended for IHC. We have also generated a mouse monoclonal and a rabbit polyclonal antibody to the same ACIII peptide, MCA-1A12 and RPCA-ACIII.
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| Name: | Chicken Polyclonal Antibody to Adenylate Cyclase III |
| Immunogen: | C-terminal peptide of rat ACIII, PAAFPNGSSVTLPHQVVDNP with a Cys added to the N-terminus to allow coupling to KLH. |
| HGNC Name: | ADCY3 |
| UniProt: | P21932 |
| Molecular Weight: | ~120kDa and above |
| Host: | Chicken |
| Species Cross-Reactivity: | Rat, Mouse, Human |
| RRID: | AB_2744500 |
| Format: | Affinity purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3 |
| Applications: | WB, IF/ICC |
| Recommended Dilutions: | WB: 1:500-1:1,000, IF/ICC 1:5,000-1:10,000, IHC not recommended |
| Storage: | Store at 4°C for short term, and at -20°C for longer term. |
G protein coupled receptors are a large and variable family of membrane proteins. On binding their specific ligand they activate specific members of the family of trimeric G-proteins which in turn activate other signalling enzymes including adenylate cyclases. Adenylate cyclases are activated by the GTP bound GαS subunits of trimeric G-proteins and are responsible for the production of the important “second messenger” signaling molecule cyclic-AMP which in turn activates the cAMP dependent protein kinase. Several different adenylate cyclase genes produce protein products with distinctly differen expression patterns in cells and tissues. The type III adenylate cyclase enzyme is specifically localized in the membranes surrounding neuronal cilia, and is activated by specific G-protein coupled receptors also located in cilia (1-5). Neuronal cilia express a variety of other receptors types and mediators of other signaling pathways and appear to function as a unique and complex neuronal sensory structure (1-5). For examples, the somatostatin 3 receptor, neuropeptide Y2 receptor and melanin concentrating hormone receptor 1 are localized in neuronal cilia and the sonic hedgehog and Wnt signalling pathway act on neurons primarily through neuronal cilia (6).
We tested this antibody for IHC on rodent and human brain sections and as a result do not recommended it for that purpose. However we found that another EnCor antibody, RPCA-ACIII, did work well for IHC, see image below. Mouse select image for larger view.
1. Fuchs JL, Schwark HD. Neuronal primary cilia: a review. Cell Biol. Int. 28:111-8 (2004).
2. Louvi A and Grove EA. Cilia in the CNS: the quiet organelle claims center stage. Neuron 69:1046-60 (2011).
3. Singla V, Reiter JF. The primary cilium as the cell's antenna: signaling at a sensory organelle. Science 313:629-33 (2006).
4. Green JA, Mykytyn K. Neuronal Primary Cilia: An Underappreciated Signaling and Sensory Organelle in the Brain. Neuropsychopharm. 39:244–5 (2014).
5. May-Simera HL, Kelley MW. Cilia, Wnt signaling, and the cytoskeleton. Cilia 2;1:7 (2012).
6. Guemez-Gamboa A, et al. Primary cilia in the developing and mature brain. Neuron 82:511-21 (2014).
7. Guadiana SM, et al. Arborization of Dendrites by developing neocortical neurons is dependent on primary cilia and Type 3 adenylyl cyclase. J. Neurosci. 33:2626-38 (2013).
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