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Immunofluorescence analysis of Connexin 50 was done on 90% confluent log phase CaCo-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with Connexin 50 Mouse Monoclonal Antibody (334300) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing junctional localization of Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059). and panel e is a no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse, Sheep, Rat|
|Published species reactivity||Pig, Rat, Mouse|
|Host / Isotype||Mouse / IgM, kappa|
|Immunogen||This antibody was prepared against urea/alkali stripped sheep lens membranes.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|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.
GJA8 is a an integral membrane protein that belongs to the connexin family, alpha-type (group II) subfamily. One gap junction consists of a cluster of closely packed pairs of transmembrane channels, the connexons, through which materials of low MW diffuse from one cell to a neighboring cell. A connexon is composed of a hexamer of connexins. This particular connexin only forms junctional channels. GJA8 is expressed in the eye lens, and defects in GJA8 are the cause of zonular pulverulent cataract type 1 (CZP1), a form of autosomal dominant congenital cataract.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Connexin50 couples axon terminals of mouse horizontal cells by homotypic gap junctions.
33-4300 was used in immunocytochemistry, immunohistochemistry, and western blot to investigate homotypic gap junctions of Connexin50 in axon terminals of mouse horizontal cells
|Dorgau B,Herrling R,Schultz K,Greb H,Segelken J,Ströh S,Bolte P,Weiler R,Dedek K,Janssen-Bienhold U||The Journal of comparative neurology (523:2062)||2015|
||Hyposmotic stress causes ATP release and stimulates Na,K-ATPase activity in porcine lens.||Shahidullah M,Mandal A,Beimgraben C,Delamere NA||Journal of cellular physiology (227:1428)||2012|
|Pig||Not Cited||Hyposmotic stress causes ATP release and stimulates Na,K-ATPase activity in porcine lens.||Shahidullah M,Mandal A,Beimgraben C,Delamere NA||Journal of cellular physiology (227:1428)||2012|
PKCγ, role in lens differentiation and gap junction coupling.
33-4300 was used in western blot to use knock out mice to determine the role of PKCgamma in the regulation of gap junction coupling in the normal lens.
|Das S,Wang H,Molina SA,Martinez-Wittinghan FJ,Jena S,Bossmann LK,Miller KA,Mathias RT,Takemoto DJ||Current eye research (36:620)||2011|
|Rat||Not Cited||Regulation of lens cell-to-cell communication by activation of PKCgamma and disassembly of Cx50 channels.||Zampighi GA,Planells AM,Lin D,Takemoto D||Investigative ophthalmology & visual science (46:3247)||2005|
alpha 8 connexin; cell surface glycoprotein; connexin 50; Connexin-49; Connexin-50; Cx49; Cx50; gap junction alpha 8; Gap junction alpha-8 protein; gap junction membrane channel protein alpha 8; gap junction membrane channel protein alpha-8; gap junction protein alpha 8 50kDa; gap junction protein, alpha 8, 50kDa; Lens fiber protein MP70; lens intrinsic membrane protein MP70; lens opacity 10; MP38; MP64
Aey5; CAE; CAE1; Cnx50; CTRCT1; CX50; CZP1; GJA8; Lop10; MP70