The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 23973667
Use a concentration of 2.5 µg/ml. Detects a band of approximately 33 kDa (predicted molecular weight: 32 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Use a concentration of 4 - 8 µg/ml.
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.
Involvement in disease
Defects in GJB1 are the cause of Charcot-Marie-Tooth disease X-linked type 1 (CMTX1) [MIM:302800]; also designated CMT-X. CMTX1 is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathies characterized by severely reduced motor nerve conduction velocities (NCVs) (less than 38m/s) and segmental demyelination and remyelination, and primary peripheral axonal neuropathies characterized by normal or mildly reduced NCVs and chronic axonal degeneration and regeneration on nerve biopsy. CMTX1 has both demyelinating and axonal features. Central nervous system involvement may occur. Defects in GJB1 may contribute to the phenotype of Dejerine-Sottas syndrome (DSS) [MIM:145900]; also known as Dejerine-Sottas neuropathy (DSN) or hereditary motor and sensory neuropathy III (HMSN3). DSS is a severe degenerating neuropathy of the demyelinating Charcot-Marie-Tooth disease category, with onset by age 2 years. DSS is characterized by motor and sensory neuropathy with very slow nerve conduction velocities, increased cerebrospinal fluid protein concentrations, hypertrophic nerve changes, delayed age of walking as well as areflexia. There are both autosomal dominant and autosomal recessive forms of Dejerine-Sottas syndrome.
Belongs to the connexin family. Beta-type (group I) subfamily.
Immunocytochemistry/ Immunofluorescence - Anti-Connexin 32 / GJB1 antibody (ab66613)Image from Ravasi M et al., Exp Cell Res. 2013;319(19):2989-99. Fig 2.; doi: 10.1016/j.yexcr.2013.08.016 with permission from Elsevier.
Immunocytochemistry/Immunofluorescence analysis of rat neuron cells co-cultured with rat MSCs labelling Connexin 32 / GLB1 wiith ab66613 at 1/50. Cultures were fixed with 4% paraformaldehyde. Green - Cx32, Red - Phalloidin, Blue - MAP2, Propidium Iodide - nuclei. Scale Bar 25 µm.
After 30 days of culture, Connexin 32 was present only in co-cultured neurons but absent in neurons cultured alone and in MSCs.
Confocal analysis of Z axis revealed that Connexin 32 was adherent to the axons.
Western blot - Connexin 32 / GJB1 antibody (ab66613)
Anti-Connexin 32 / GJB1 antibody (ab66613) at 2.5 µg/ml + Jurkat cell lysate at 10 µg
Secondary HRP conjugated anti-rabbit IgG at 1/50000 dilution
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human intestine tissue labelling Connexin 32 / GJB1 with ab66613 at 4-8µg/ml. Arrows indicate positively labelled epithelial cells of the intestinal villus. Magnification: 400X.
Kim S et al. The basic helix-loop-helix transcription factor E47 reprograms human pancreatic cancer cells to a quiescent acinar state with reduced tumorigenic potential. Pancreas44:718-27 (2015).
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Ravasi M et al. Undifferentiated MSCs are able to myelinate DRG neuron processes through p75. Exp Cell Res319:2989-99 (2013).
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