The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).
Essential for bone resorption and osteoclast differentiation (By similarity). Reversible hydration of carbon dioxide. Can hydrates cyanamide to urea. Involved in the regulation of fluid secretion into the anterior chamber of the eye.
Defects in CA2 are the cause of osteopetrosis autosomal recessive type 3 (OPTB3) [MIM:259730]; also known as osteopetrosis with renal tubular acidosis, carbonic anhydrase II deficiency syndrome, Guibaud-Vainsel syndrome or marble brain disease. Osteopetrosis is a rare genetic disease characterized by abnormally dense bone, due to defective resorption of immature bone. The disorder occurs in two forms: a severe autosomal recessive form occurring in utero, infancy, or childhood, and a benign autosomal dominant form occurring in adolescence or adulthood. Autosomal recessive osteopetrosis is usually associated with normal or elevated amount of non-functional osteoclasts. OPTB3 is associated with renal tubular acidosis, cerebral calcification (marble brain disease) and in some cases with mental retardation.
Western blot - Anti-Carbonic Anhydrase II antibody (ab82559)
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: Carbonic Anhydrase II knockout HAP1 cell lysate (20 µg) Lane 3: A431 cell lysate (20 µg) Lane 4: HEk293 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab82559 observed at 30 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab82559 was shown to specifically react with Carbonic Anhydrase II when Carbonic Anhydrase II knockout samples were used. Wild-type and Carbonic Anhydrase II knockout samples were subjected to SDS-PAGE. ab82559 and ab8245 (loading control to GAPDH) were diluted at 1 μg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Carbonic Anhydrase II antibody (ab82559)
All lanes : Anti-Carbonic Anhydrase II antibody (ab82559) at 1 µg/ml
Lane 1 : Bone Marrow (Human) Tissue Lysate - adult normal tissue Lane 2 : Human small intestine tissue lysate - total protein (ab29276) Lane 3 : Human kidney tissue lysate - total protein (ab30203) Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 29 kDa Observed band size: 29 kDa