SH‐SY cells grown on coverslips coated with Poly‐L lysine were fixed with 4%PFA (Electron Microscopy Sciences) for 15 min and then washed in D‐PBS. Cells were permeabilized with D‐PBS/0.2% Triton X‐100 for 1 h, then blocked in 3% Normal Donkey serum in D‐PBS/0.2% Triton X‐100 for 1 h. Next, cells were incubated O.N. at 4c with 1:100 dilution of anti‐NCX1 antibody (Abcam—ab2869) in blocking solution. Cells were washed in D‐PBS/0.2% Triton X‐100 and incubated for 1 h at RT with 1:500 dilution of goat anti‐mouse Cy3‐conjugated secondary antibody (Jackson immunoresearch 115‐166‐072). Finally, cells were washed in D‐PBS, stained with DAPI for 10 min, washed, and mounted on microscope slides.
Anti ncx1 antibody
Manufactured by Abcam
Anti-NCX1 antibody is a primary antibody that recognizes the Na+/Ca2+ exchanger 1 (NCX1) protein. NCX1 is an integral membrane protein that facilitates the exchange of sodium and calcium ions across the cell membrane. This antibody can be used to detect and study the expression and localization of NCX1 in various cell and tissue types.
Automatically generated - may contain errors
Lab products found in correlation
Cy3 conjugated goat anti mouse secondary antibody, by Jackson ImmunoResearch (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Inhibitor bay 11 7082, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Kb r7943, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
2 protocols using anti ncx1 antibody
1
Immunocytochemistry for NCX1 in SH-SY Cells
2
Pancreatic Acinar Cell NF-κB Regulation
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The commercially available reagents cerulein (MCE, Shanghai, China), l -arginine (MCE), NCX1 inhibitor KB-R7943 (Sigma-Aldrich, St. Louis, Mo), nuclear factor-κB (NF-κB) inhibitor Bay 11–7082 (Sigma-Aldrich), and NF-κB activator Phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich) were all purchased through the indicated manufacturers. Anti-NCX1 antibody was produced by Abcam (Cambridge, UK). Antibodies against P65, pP65(ser536), GAPDH, β-actin, and goat anti-rabbit/mouse IgG (H + L) secondary antibodies were purchased from Cell Signaling Technologies (Danvers, Mass). The rat pancreatic acinar cell line AR42J and human embryonic kidney cell line HEK-293T were both supplied by the Cell Line Resource Center of the Chinese Academy of Medical Sciences (Beijing, China). All cell lines were stored frozen in liquid nitrogen and after thawing, less than 20 of passages were used over the course of 3 months for the present experiments.
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