Anti-CARMA/CARD *NEW* IHC Validated


AnaSpec is excited to announce the release of a series of newly IHC validated antibodies. This month, we introduce the CARMA/CARD/Bimp antibodies.

CARMA proteins are scaffold molecules that belong to the membrane-associated guanylate kinase-like (MAGUK) and the caspase-associated recruitment domain (CARD) protein families. They play critical role in antigen receptor signaling pathways.1-4 CARMA-1 (CARD-11) CARMA-2 (CARD-14), and CARMA-3 (CARD-10) share homologous sequence and functions but are found in different tissues. CARMA1 is predominantly located in lymphoid tissue.5 CARMA-1 protein associates with lipid rafts1 and interacts with Bcl10, that is required for activation of TCR induced NFkB signaling pathway.1,2,4 Specifically, CARMA-1 N-terminal CARD domain binds to Bcl10 CARD motif.5 It is also essential for protein-induced IL-2 and INF-g production.2 CARMA-2 is mostly found in placenta.5 CARMA-2 protein induces NFkB activation; however, its signaling pathway it is not yet known.5 CARMA-3 is most abundant in nonhematopoietic cells.5 CARMA-3 protein physically associates with IKK complex and induces NFkB activation via LPA signaling pathway.7 CARMA genes deactivation studies in mice resulted in reduced immunoglobulin levels and absence of TCR-induced NFkB activation.4,6,7 This suggests that CARMA mutations may lead to adaptive and innate immunity malfunction.

Immunohistochemistry of these antibodies was performed by an anatomic pathologist on formalin-fixed, paraffin-embedded human or mouse tissues. Table 1 summarizes the IHC results. At least 22 different human or mouse tissues or cell types, including positive and negative tissues were used for each antibody tested. Dilutions of the antibodies at 2.5, 5, 10 and 20 ug/ml were tested, with the optimal concentration found to be at 10 ug/ml. These antibodies, provided at 50 ug, are sufficient for at least 5 IHCs. Moderate to strong staining with minimal background staining at the expected target tissues were observed for the antibodies. A representative image from a positively stained tissue is shown for each antibody. It shows the localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.

Table 1. IHC results of the CARMA antibodies.

Staining Intensity

Tissues Stained

IHC

Anti-CARMA-1/CARD-11, human, Cat# 54982N

Moderate to strong

Human lymphoid organs: tonsil, lymph node, thymus, and spleen

Anti-CARMA-1/CARD-11, mouse, Cat# 54985N

Moderate to strong

Mouse lymphoid organs: tonsil, lymph node, thymus, and spleen

Anti-CARMA-2/CARD-14,human, Cat# 54983N

Moderate

Variety of human cell types, including trophoblasts of the placenta

Anti-CARMA-3/CARD-10,human, Cat# 54984N

Moderate

Human tissues: heart, kidney, small and large intestine, skeletal muscle, and testis

Anti-CARMA-3/CARD-10, mouse, Cat# 54987N

Moderate

Mouse tissues: heart, intestine, skeletal muscle, and testis

REFERENCES

1. Gaide, O et al. Nat. Immunology 3: 836 (2002).
2. Hara, H. et al. JEM, 200, 1167 (2004).
3. Narayan, P. et al. Mol Cell Bio 26, 2327 (2006).
4. Egawa T. et al. Current Biology 13, 1252 (2003).
5. Grabiner, B. et al. Genes and Dev. 21, 984 (2007).
6. Newton, K. et al. Current Biology 13, 1247 (2003).
7. Stilo R. et al. J. Biol. Chem.279: 34323 (2004).