The family of EF-hand type Ca2+-binding proteins includes Calbindin (previously designated vitamin D-dependent Ca2+-binding protein), S-100 α and β, Cal-granulins A (also designated MRP8), B (also designated MRP14) and C (S-100 like proteins) and the parvalbumin family members, including parvalbumin α and parvalbumin β (also designated oncomodulin). Calbindin, S-100 proteins and parvalbumin proteins are each expressed in neural tissues. In addition, S-100 α and β are present in a variety of other tissues and Calbindin is present in intestine and kidney. Parvalbumin α is also found in fast-contracting/ relaxing skeletal muscle fibers and parvalbumin β is found in many tumor tissues as well as in the organ of Corti. Calbindin, S-100 proteins and parvalbulmins have all been detected in Leydig cells and the testis. These proteins are thought to play a role in hormone production and spermatogenesis. Calgranulin is expressed in macrophages and epithelial cells.
Background References
1. Rebouillat D. et. al. The 100-kDa 2\',5\'-oligoadenylate synthetase catalyzing preferentially the synthesis of dimeric pppA2\'p5\'A molecules is composed of three homologous domains. J. Biol. Chem. 274:1557-1565(1999).
Western blot analysis of OAS3 on different lysates with Rabbit anti-OAS3 antibody (HA500039) at 1/1,000 dilution.
Lane 1: Mouse testis tissue lysate Lane 2: Rat testis tissue lysate
Lysates/proteins at 40 µg/Lane.
Predicted band size: 126 kDa Observed band size: 126 kDa
Exposure time: 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500039) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-OAS3 antibody (HA500039) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500039) at 1/1,000 dilution for 1 hour 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.
Immunocytochemistry analysis of A594 cells labeling OAS3 with Rabbit anti-OAS3 antibody (HA500039) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-OAS3 antibody (HA500039) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Immunocytochemistry analysis of LOVO cells labeling OAS3 with Rabbit anti-OAS3 antibody (HA500039) at 1/200 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-OAS3 antibody (HA500039) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
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