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||Rabbit, Pig, Rat, Mouse, Human, Not Applicable|
|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.
|Western Blot (WB)||See 6 publications below|
|Miscellaneous PubMed (MISC)||See 1 publications below|
|Immunocytochemistry (ICC)||See 1 publications below|
|Immunofluorescence (IF)||See 1 publications below|
|Immunohistochemistry (Paraffin) (IHC (P))||See 1 publications below|
|Immunohistochemistry (Frozen) (IHC (F))||See 1 publications below|
|Immunoprecipitation (IP)||See 1 publications below|
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.
Connexin 50 modulates Sox2 expression in spinal-cord-derived ependymal stem/progenitor cells.
33-4300 was used in immunocytochemistry and western blot to investigate modulation of Sox2 expression in spinal-cord-derived ependymal stem/progenitor cells by connexin 50
|Rodriguez-Jimenez FJ,Alastrue A,Stojkovic M,Erceg S,Moreno-Manzano V||Cell and tissue research (365:295)||2016|
|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|
PKC¿, 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 and visual science (46:3247)||2005|
|Rat||Not Cited||Differential phosphorylation of connexin46 and connexin50 by H2O2 activation of protein kinase Cgamma.||Lin D,Lobell S,Jewell A,Takemoto DJ||Molecular vision (10:688)||2004|
|Growth factors but not gap junctions play a role in injury-induced Ca2+ waves in epithelial cells.||Klepeis VE,Cornell-Bell A,Trinkaus-Randall V||Journal of cell science (114:4185)||2001|
Expression and Localization of Connexins in the Outer Retina of the Mouse.
33-4300 was used in immunohistochemistry (frozen) to investigate the counterpart of connexin36 at gap junctions between rods and cones.
|Bolte P,Herrling R,Dorgau B,Schultz K,Feigenspan A,Weiler R,Dedek K,Janssen-Bienhold U||Journal of molecular neuroscience : MN (58:178)||2016|
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|
Expression of connexins in chromaffin cells of normal human adrenals and in benign and malignant pheochromocytomas.
33-4300 was used in immunohistochemistry - paraffin section to examine connexin expression in chromaffin cells of normal human adrenal glands and benign and malignant pheochromocytomas
|Willenberg HS,Schott M,Saeger W,Tries A,Scherbaum WA,Bornstein SR||Annals of the New York Academy of Sciences (1073:578)||2006|
|Not Applicable||5-10 µg/ml||
Gap junctional coupling and connexin immunoreactivity in rabbit retinal glia.
33-4300 was used in immunohistochemistry - frozen section to study connexins in mammalian Muller cells
|Zahs KR,Ceelen PW||Visual neuroscience (23:1)||2006|
||Differential phosphorylation of connexin46 and connexin50 by H2O2 activation of protein kinase Cgamma.||Lin D,Lobell S,Jewell A,Takemoto DJ||Molecular vision (10:688)||2004|