Immunofluorescence analysis of Cx46 was performed on cryostat sections of adult mouse lens. Tissue sections on slides were probed for 24 h at 4°C in a humidified chamber with rabbit polyclonal anti-Cx46 (Cat. No. 38-8300) at an antibody concentration of 2 µg/ml diluted in 50 mM Tris-HCl, pH 7.4, containing 1.5% NaCl, 0.3% Triton X-100 (TBST) and 4% normal goat serum. After overnight incubation, sections were washed extensively for 1 h in TBST, and detection of primary antibody was performed for 1.5 h at room temperature with AlexaFluor-488-conjugated donkey anti-rabbit diluted 1:600 in TBST. Sections were then washed in TBST, then in TBS (without triton) and then coversliped with anti-fade medium. Images were taken on a Zeiss 710 confocal microscope at x40 objective magnification, and show immunofluorescence labelling of Cx46 in gap junctions between cells in mouse lens.
|Tested species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from an internal region of the mouse Connexin 46 protein|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||2 µg/ml|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
38-8300 detects Cx46 in IHC application in cryostat sections of adult mouse lens.
Connexin 46 (Cx46), also known as Gap Junction Protein Alpha-3 (GJA3), CAE3, CX46 and C2P3, maps to human chromosome 13q11-q12 and encodes a 46 kDa protein. Cx46, along with Cx50, is principally expressed in the lens of the eye. Cx46 mediates intercellular interactions during development and is necessary for the survival of neural crest cells. Cx46 forms gap junctions that connect lens fiber cells and are crucial for maintaining lens transparency and normal lens function. Mutations of Cx46 result in severe cataracts of the lens. Individual knockouts of Cx46 and Cx50 lead to changes in the rate of lens fiber cell differentiation and cell size, thus the interaction of Cx46 and Cx50 is required for proper organization of fiber cells.
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