The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Predicted molecular weight: 49.7 kDa.
1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 5 µg/ml.
FunctionReversible hydration of carbon dioxide. Participates in pH regulation. May be involved in the control of cell proliferation and transformation. Appears to be a novel specific biomarker for a cervical neoplasia.
Tissue specificityExpressed primarily in carcinoma cells lines. Expression is restricted to very few normal tissues and the most abundant expression is found in the epithelial cells of gastric mucosa.
Sequence similaritiesBelongs to the alpha-carbonic anhydrase family.
Post-translational modificationsAsn-346 bears high-mannose type glycan structures.
Cellular localizationNucleus. Nucleus > nucleolus. Cell membrane. Cell projection > microvillus membrane. Found on the surface microvilli and in the nucleus, particularly in nucleolus.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Carbonic Anhydrase IX antibody (ab10471)This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.
ab10471 staining Carbonic Anhydrase IX in Human kidney tissue sections by IHC-P (Formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5 minutes of peroxidase block followed by 10 minutes of protein block at 20°C; antigen retrieval was heat mediated in retrieval solution. Samples were incubated with primary antibody (1/250 in antibody diluent) for 45 minutes at 20°C. An undiluted HRP-conjugated polymer goat anti-mouse/rabbit IgG polyclonal was used as secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Carbonic Anhydrase IX antibody (ab10471)
ICC/IF image of ab10471 stained DU145 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab10471 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-Carbonic Anhydrase IX antibody (ab10471)
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