The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. See Abreview.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/2000. Predicted molecular weight: 60 kDa.
FunctionOccurs in almost all aerobically respiring organisms and serves to protect cells from the toxic effects of hydrogen peroxide. Promotes growth of cells including T-cells, B-cells, myeloid leukemia cells, melanoma cells, mastocytoma cells and normal and transformed fibroblast cells.
Involvement in diseaseDefects in CAT are the cause of acatalasia (ACATLAS) [MIM:115500]; also known as acatalasemia. This disease is characterized by absence of catalase activity in red cells and is often associated with ulcerating oral lesions.
Sequence similaritiesBelongs to the catalase family.
Post-translational modificationsThe N-terminus is blocked.
ICC/IF image of ab16731 stained Hela cells. The cells were 4% PFA fixed (10 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 (ab16731, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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.
Ab16731 staining human normal adrenal gland tissue. Staining is localised to intracellular compartment (peroxisomes). Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification
Western blot - Anti-Catalase antibody (ab16731)
All lanes : Anti-Catalase antibody (ab16731) at 1/2000 dilution
Lane 1 : 40ug supernatant of mouse liver homogenate Lane 2 : 20ug supernatant of mouse liver homogenate Lane 3 : 5ug supernatant of mouse liver homogenate
Secondary HRP conjugated donkey anti-rabbit antibody Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 60 kDa Observed band size : 60 kDa
Exposure time : 1 minuteThis image is courtesy of an Abreview submitted by Sandra Sobocanec on 16 March 2006.
Immunocytochemistry/ Immunofluorescence - Anti-Catalase antibody (ab16731)This image is a courtesy of an anonymous Abreview.
ab16731 at 1/200 dilution staining Catalase in human 293FT cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde and blocked in 5% BSA for 1 hour at 25°C. The primary antibody was used at 1/200 dilution in PBS and incubated with sample at 4°C for 12 hours. An Alexa Fluor® 488 conjugated Goat polyclonal to rabbit IgG was used undiluted as secondary.
Immunocytochemistry/ Immunofluorescence - Anti-Catalase antibody (ab16731)This image is courtesy of an anonymous abreview.
ab16731 at a 1/200 dilution staining Catalase in mouse bone marrow cells by Immunocytochemistry/ Immunofluorescence, incubated for 9 hours at 4°C. Formalin fixed. Blocked with 2% BSA for 30 minutes at 20°C. Secondary used at 1/200 dilution polyclonal Goat anti-rabbit IgG conjugated to Alexa Fluor 488 (green). Nuclei stained with DAPI (blue).
Immunohistochemistry (Frozen sections) - Anti-Catalase antibody (ab16731)This image is a courtesy of Anonymous Abreview
ab16731 staining Catalase in mouse liver tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and permeabilized with 0.2% Triton X-100 before blocking with 2% BSA for 30 minutes at 200C. The sample was incubated with primary antibody (1/200) for 9 hours at 40C. An Alexa Fluor®488-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution. DAPI was used to stain the cell nuclei (blue).
Schiffer TA et al. Control of human energy expenditure by cytochrome c oxidase subunit IV-2. Am J Physiol Cell Physiol311:C452-61 (2016).
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Liu X et al. The novel triterpenoid RTA 408 protects human retinal pigment epithelial cells against H2O2-induced cell injury via NF-E2-related factor 2 (Nrf2) activation. Redox Biol8:98-109 (2016).
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