Technical Data
CLAN (NLR Family CARD Domain-containing Protein 4, CARD, LRR, and NACHT-containing Protein, Clan Protein, Caspase Recruitment Domain-containing Protein 12, Ice Protease-activating Factor, Ipaf, NLRC4, CARD12, CLAN1, IPAF, UNQ6189/PRO20215)
Ipaf (also known as Clan/CARD12) is a CARD domain containing protein. CARD (caspase-associated recruitment domain) proteins are key regulators of cell death, cell survival and cytokine production. In general CARD proteins are implicated in host defense against infection, environmental stress or cellular damage. CARD domains are found in the N-terminal pro-domains of certain caspases, a family of apoptotic and pro-inflammatory proteases, as well as in a diversity of other proteins including Ipaf/Clan/CARD12. CARD domains are homotypic protein interaction motifs that enable networks of proteins to communicate via CARD-CARD interactions. There are at least three major signaling pathways in which CARD proteins act: (1) Regulation of caspase activation in the context of apoptosis (2) Regulation of caspase activation in the context of inflammation (3) Regaultion of NF-kB activation in the context of innate or adaptive immune responses. As there is significant crosstalk between pathways that lead to caspase-mediated apoptosis or inflammation and pathways that result in NF-kB activation, it is logical that similar protein modules such as CARD domains are found repeatedly in proteins from all three pathways. Ipaf plays a role in regulating caspase-1 activity, which in turn mediates the maturation of inflammatory cytokines IL-1b and IL-18. In transfected cells, Ipaf has been shown to directly interact with procaspase-1 and induce proteolytic activation of procaspase-1 in transfected cells. On the flip side, macrophages from IPAF deficient mice failed to activate caspase-1 in response to Salmonella typhimurium infection underscoring the importance of IPAF in vivo. IPAF also interact with the pro-apoptotic adaptor protein ASC and co-expression of IPAF with ASC has been shown to induce NF-kB activation and apoptosis.

Suitable for use in Western Blot, Immunohistochemistry and Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:2000
Immunohistochemistry (formalin fixed paraffin embedded): 1:1000-1:5000
Immunoprecipitation: 1:50-1:200
Immunohistochemistry: Frozen
Optimal dilutions to be determined by the researcher.

Positive Control:
Spleen, lymphoid, many cancer cell lines

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
50ul-20CBlue IceHumanRabbit
Not determined.
Synthetic peptide corresponding to aa637-654 (EEAPETYIPSRAVSLFFN) of CLAN; GenBank no. gi|40788015|ref|NP_067032.3|.
Supplied as a liquid, 0.05% sodium azide.
Recognizes CLAN. Species Crossreactivity: mouse and human. Species sequence homology: rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Damiano JS and JC Reed. 2004. CARD proteins as therapeutic targets in cancer. Current Drug Targets 5:367-374. 2. Lu C, A Wang, L Wang, M Dorsch, TD Ocain, Y Xu. 2005. Nucleotide binding to CARD12 and its role in CARD12-mediated caspase-1 activation. Biochemical and Biophysical Research Communications 331:1114-1119.