Eosinophil cationic protein (ECP) also known as ribonuclease 3 is a basic protein located in the eosinophil primary matrix. In humans, the eosinophil cationic protein is encoded by the RNASE3 gene. ECP is released during degranulation of eosinophils. This protein is related to inflammation and asthma because in these cases, there are increased levels of ECP in the body. There are three glycosolated forms of ECP and consequently ECP has a range of molecular weights from 18-22 kDa. Eosinophil cationic protein and the sequence related eosinophil-derived neurotoxin (RNASE2) are both members of the Ribonuclease A superfamily. Both proteins possess neurotoxic, helmintho-toxic, and ribonucleo-lytic activities. Eosinophil cationic protein is localized to the granule matrix of the eosinophil. Eosinophil granulocytes appear in large numbers in inflammation sites and in response to certain parasitic infections. These cytoplasmic granules contain positively charged proteins that characterize the cells. ECP is one of the four highly basic proteins that enter the surrounding tissues when activated eosinophils degranulate. There are several mechanisms that can be combined to generate an asthma attack, including specific IgE antibodies, activated inflammatory cells, neurogenic mechanisms, hyperresponsiveness and individual hormonal imbalances. Allergic reactions in the lung typically have two phases. The late phase typically occurs several hours after exposure, upon which eosinophils accumulate in the bronchus and release granule proteins that cause bronchial irritability.
Background References
1. Diop G. et. al. Genetic variants of RNASE3 (ECP) and susceptibility to severe malaria in Senegalese population. Malar J. 2018 Feb
2. Geng R. et. al. Differential Expression of Six Rnase2 and Three Rnase3 Paralogs Identified in Blunt Snout Bream in Response to Aeromonas hydrophila Infection. Genes (Basel). 2018 Feb
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Ribonuclease 3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (R1511-19, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Ribonuclease 3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (R1511-19, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western blot analysis of Ribonuclease 3 on different lysates with Rabbit anti-Ribonuclease 3 antibody (R1511-19) at 1/1,000 dilution.
Lane 1: U937 (Human histiocytic lymphoma cells) cell lysate
Exposure time: 1 minute 50seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: R1511-19, 1/1,000 in primary antibody dilution buffer 5% NFDM/TBST, overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 18.4 kDa Observed band size: 18 kDa
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