Calcium Sensing Receptor Antibody (5C10, ADD)

Immunohistochemistry was performed on normal biopsies of deparaffinized Human brain tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Calcium Sensing Receptor (MA1-934) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on normal biopsies of deparaffinized Human kidney tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Calcium Sensing Receptor (MA1-934) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemical analysis of mouse stomach stained with Calcium Sensing Receptor Monoclonal Antibody (Product # MA1-934). Fresh stomach tissue was fixed in 4% formalin for 1 hour and then incubated overnight at 4ºC in 25% sucrose before embedding in tissue freezing medium. Antigen retrieval was carried out on 8µm cryo-sections by incubating in sodium citrate buffer for 45 minutes at 4ºC, immersing in sodium citrate buffer for 10 minutes at 100ºC before washing 3 times for 5 minutes each in 1X PBS. Sections were then blocked in blocking buffer (0.3% Triton X-100 in 1X PBS containing 10% normal goat serum) for 30 minutes at RT before staining with MA1-934 (diluted 1:100 in blocking buffer) overnight at 4ºC followed by a fluorophore-conjugated goat anti-mouse IgG secondary antibody for 2 hours at RT. Sections were also stained with DAPI nuclear stain (blue). MA1-934 positive cells (green) were found at the base of the antral glands in the mouse stomach. Data courtesy of the Innovators Program.