Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party scientific content. Neither the content nor the BenchSci technology and processes for selection have been evaluated by us; we are providing them as-is and without warranty of any kind, including for use or application of the Thermo Fisher Scientific products presented.    Immunofluorescence analysis of Connexin47 (Cx47) was performed on sections of adult mouse brain. Tissue sections on slides were probed for 24 h at 4°C in a humidified chamber with rabbit polyclonal anti-Cx47 (Product # 36-4700) at an antibody concentration of 1-2 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 Z2 scanning microscope at x40 objective magnification, and show immunofluorescence labelling of Cx47 localized at gap junctions between astrocyte processes and the cell bodies and initial processes of oligodendrocytes in the thalamus of adult mouse brain. Data courtesy of Dr. James Nagy\'s lab.   Immunohistochemistry analysis of Connexin 47 showing staining in the membrane and cytoplasm of paraffin-embedded mouse cerebellum tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Connexin 47 polyclonal antibody (Product # 36-4700) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.   Immunohistochemistry analysis of Connexin 47 showing staining in the membrane and cytoplasm of paraffin-embedded human brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Connexin 47 polyclonal antibody (Product # 36-4700) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Immunohistochemistry   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Immunocytochemistry   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Western Blot   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Immunohistochemistry   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Immunohistochemistry   Figure 2 mCx47 M282T and beta-gal expression in oligodendrocytes of 40-day-old mutant mice. A-C, LacZ staining of 50 um brain slices obtained from Cx47 +/M282T mice. The beta-gal activity is restricted to nuclear localisation. A, beta-Gal positive cells in the corpus callosum show typical oligodendrocytic chain-like organization and coexpression of the oligodendrocytic marker CNPase. LacZ expression was not detectable in GFAP-positive astrocytes (B) or NeuN-positive neurons (C). D, Immunostaining with Cx47 antibodies (green) revealed signals in close proximity of Syto-61 stained nuclei in the corpus callosum of Cx47 +/+ mice. Weaker signals were also found more distal to the nuclei. E, Cx47 antibody stainings on heterozygous Cx47 +/M282T mice resulted in weaker, but similarly localized immunosignals compared to those obtained on wildtype brain tissue. Cx47 gap junction immunosignals were predominantly localized close to beta-gal positive nuclei indicated by antibody staining. F, Homozygous Cx47 M282T/M282T mice showed apparent beta-gal immunoreactivity, but robust Cx47 gap junction immunosignals were not detected in the perikarya. Very weak Cx47 signals were noticed in brain tissue of Cx47 M282T/M282T mice. G, Immunoblot staining against beta-gal on whole brain lysates of 40-day-old mice yielded strong signals with Cx47 M282T/M282T , weaker signals with Cx47 +/M282T and no signals with Cx47 +/+ tissue. H, Immunoblot analysis on Cx47 protein resulted in unspecific bands at 50   Figure 10 Expression of mutant mCx47 M282T results in delayed oligodendrocytic differentiation. Immunoblots were performed with cerebellar lysates. At least three animals per group were analyzed. For each oligodendrocytic marker, protein levels were normalized to beta-tubulin expression as internal standard. Expression levels of Cx47 +/+ mice were set to 100% per group. A, Densitometric quantification of immunoblot analyses revealed significantly decreased CNPase expression in P10, P16 and P40 Cx47 M282T/M282T mice compared to their wildtype littermates (** p   Figure 1 Generation of mCx47 M282T mice with LacZ reporter gene. A, Scheme of homologous recombination of the targeting vector into the Cx47 coding region. The resulting transgenic allele ( Cx47M282Tneo ) includes mCx47 M282T coding DNA, an internal ribosome entry site (IRES) followed by a nuclear localization signal (nls) fused to LacZ coding DNA and a neomycin selection cassette flanked by frt-sites. The endonuclease Bsm BI restriction site was generated by T to C transition on nucleotide 845 of the Gjc2 coding region, resulting in the mutant mCx47 M282T . B, Flp recombinase activity causes deletion of the neomycin selection cassette. Specific primer binding sites and Mfe I restriction sites are indicated. C, PCR products specific for wild-type (570 bp) and transgenic loci (730 bp) using DNA obtained from tail tip tissue. D, Presence of the mutant allele was proven by PCR amplification of a 700 bp Cx47 fragment and subsequent Bsm BI digestion. Only mutant alleles yielded the 350 bp fragment. E, PCR analysis resulted in amplification of the 1,700 bp fragment of the neomycin selection cassette deprived allele and the 450 bp fragment of the Cx47M282Tneo locus. F, Homologous recombination and different allelic combinations were confirmed by Southern blot hybridization using a probe derived from the 3 region of Cx47. Mfe I digestion of genomic DNA prepared from liver resulted in the of 4.6 kb fragment for the Cx47 wildtype allele, the 10.5 kb fragment for the Cx47M282Tneo   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Immunomicroscopy   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Immunohistochemistry   Published figure using Connexin 47 polyclonal antibody (Product # 36-4700) in Western Blot Synthetic peptide derived from the C-terminal region of the mouse Connexin 47 protein. 36-4700 was used in immunofluorescence analysis of Connexin47 (Cx47) on sections of adult mouse brain. This gene encodes a member of the gap junction protein family. The gap junction proteins are membrane-spanning proteins that assemble to form gap junction channels that facilitate the transfer of ions and small molecules between cells. According to sequence similarities at the nucleotide and amino acid levels, the gap junction proteins are divided into two categories, alpha and beta. Mutations in this gene cause X-linked Charcot-Marie-Tooth disease, an inherited peripheral neuropathy. Alternatively spliced transcript variants encoding the same protein have been found for this gene. 蛋白别名: connexin 47 type A; connexin 47 type C; connexin 47 type D; Connexin-46.6; Connexin-47; Cx46.6; Cx47; CXG2; Gap junction alpha-12 protein; Gap junction gamma-2 protein; gap junction membrane channel protein alpha 12; gap junction protein, chi 2; gap junction protein, gamma 2, 47kDa Host server : magellan-srch-3-prod-green:8080/10.253.228.100:8080. git-commit: 2cd8645d2fc6bfe4ccb4abfa14772b0a94f68e98 git-url: http://victoria.invitrogen.com:8333/magellan/core.git git-branch: origin/release/1.27.0-2021.08.32-1.0