SpecificityThis antibody recognizes the 64kD polypeptide (cytokeratin 3) and 55 kDa polypeptide (cytokeratin 12) of rabbit corneal epithelium. It also recognizes bovine snout and lip epithelium, as well as a rabbit corneal cell line.
The clone number has been updated from (2Q1040) to (AE5) both clone numbers name the same antibody clone.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration. Predicted molecular weight: 54 , 65 kDa.
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
RelevanceCytokeratin 3 is a member of the intermediate filament family of proteins and is a heterotetramer of two type I and two type II keratins. Keratin 3 is specifically expressed in the corneal epithelium with family member KRT12.
Cytokeratin 12 encodes the type I intermediate filament chain keratin 12, expressed in corneal epithelia.
Defects in KRT3 and KRT12 are a cause of Meesmann corneal dystrophy (MCD), an autosomal dominant disease that causes fragility of the anterior corneal epithelium. Symptoms occur in adulthood and include rupture of the corneal microcysts that may lead to photophobia, contact lens intolerance and intermittent diminution of visual acuity. Defects in KRT12 are a cause of juvenile epithelial corneal dystrophy of Meesmann (MCD)
Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 3+12 antibody [AE5] (ab68260)This image is courtesy of an Abreview submitted by Dr Aaron Gardner
Immunofluorescence analysis of rabbit cornea cells, staining Cytokeratin 3+12 with ab68620.
Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 3% BSA for 1 hour at 20°C. Samples were incubated with primary antibody (1/50 in 3% BSA in PBST (0.5% Tween 20)) for 12 hours at 4°C. An AlexaFluor®594-conjugated goat anti-mouse polyclonal IgG (1/100) was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 3+12 antibody [AE5] (ab68260)Image cropped from Jing L et at., Molecular Vision, 17, 2263-2271, Fig. 2e
Immunofluorescent analysis of human ocular surface squamous cell carcinoma tissue, labeling Cytokeratin 3+12 with ab68260. Samples were fixed in 4% paraformaldehyde and blocked wuth 4% BSA with 0.1% Triton X-100. Incubation with ab68260 (diluted 1/50) overnight at 4°C. Samples were then incubated with an Alexa Fluor 488-conjugated secondary antibody for 1 hour, and counterstained with DAPI to visualize nuclei.
References for Anti-Cytokeratin 3+12 antibody [AE5] (ab68260)
Li J et al. S100A expression in normal corneal-limbal epithelial cells and ocular surface squamous cell carcinoma tissue. Mol Vis17:2263-71 (2011).
IHC (PFA fixed)
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