Anti cx43

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-Cx43 is a laboratory reagent that can be used to detect the presence and distribution of Connexin 43 (Cx43), a gap junction protein, in cells and tissues. It is intended for research use only.

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Lab products found in correlation

Lsm800 confocal microscope, by Zeiss (2 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Protein a g agarose bead, by Santa Cruz Biotechnology (2 mentions) Jetprime reagent, by Polyplus Transfection (2 mentions) Anti cx26, by Thermo Fisher Scientific (1 mentions) Anti cytokeratin 16, by Abcam (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Anti ki67, by Abcam (1 mentions) Alexa 594, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Zen 2 blue edition software, by Zeiss (1 mentions) Alexa fluor 555 donkey anti mouse secondary antibody, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions) C2 confocal microscope, by Nikon (1 mentions) Anti ha, by Merck Group (1 mentions) Anti aqp4, by Merck Group (1 mentions) Anti n cadherin, by Thermo Fisher Scientific (1 mentions) Mouse anti gapdh, by Merck Group (1 mentions) Anti zo 1, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Rabbit anti-Cx43 (4 citations) Mouse anti-Cx43 (4 citations) Monoclonal anti-unphosphorylated Cx43 antibody (2 citations) Mouse anti-Cx43 monoclonal antibody (2 citations)

6 protocols using anti cx43

Immunofluorescence Analysis of Cell Junctions

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Tissue taken directly from the DMEM media (~0.25 mm²) was mounted in optimal cutting temperature compound (OCT) (ThermoFisher, Paisley, UK) and 7 μm sections cut onto pre-coated slides. Tissue sections, organotypic cell models or cells cultured on 16 mm² glass coverslips were fixed in ice-cold methanol prior to permeabilisation and blocking, as previously described [14 (link)]. The primary antibodies: rabbit polyclonal anti-Cx43 (1:100 dilution, [68 (link)]); mouse monoclonal anti-Cx26 (1:50 dilution, 13–8100, ThermoFisher, Paisley, UK); rabbit polyclonal anti-Ki67 (1:500, Abcam, Cambridge, UK, ab15580); rabbit polyclonal anti-cytokeratin16 (1:100; Abcam, Cambridge, UK ab53117); rabbit polyclonal anti-Cx43^pSer368 (1:100; Sigma-Aldrich, Irvine, UK) and relevant secondary antibodies (goat anti-mouse Alexa 488 or -rabbit Alexa 594 (1:500 dilution; ThermoFisher, Paisley, UK) were used. Nuclei were counterstained with DAPI (2.5 μg/mL) (Sigma-Aldrich, Irvine, UK). Samples were visualised on a Zeiss LSM 800 confocal microscope. The mean fluorescence intensity/μm² (MFI) was extracted for repeat areas of each image captured using Zen 2. Blue edition software (Carl Zeiss Microscopy GmbH, Jena, Germany).

O’Shaughnessy E.M., Duffy W., Garcia-Vega L., Hussey K., Burden A.D., Zamiri M, & Martin P.E. (2021). Dysregulation of Connexin Expression Plays a Pivotal Role in Psoriasis. International Journal of Molecular Sciences, 22(11), 6060.

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Immunofluorescence Analysis of Connexin 43 in Immature COCs

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Paraffin-embedded sections of immature COCs were processed as previously described (31 (link)). A group of 50–60 immature COCs was collected, fixed with 4% paraformaldehyde (Solarbio, P1110) at 4°C for 12 h, and subsequently processed. After fixation, the COCs were dehydrated in 70%, 95%, and 100% alcohol at room temperature for 30 min, followed by incubation in xylene for 30 min. The COCs were then embedded in soft wax and hard wax, respectively. These paraffin blocks were cut into 5-µm thick sections, blocked with 5% normal donkey serum (Jackson ImmunoResearch Laboratories, 017-000-121), and incubated with anti-CX43 (1:100, Thermo Fisher, monoclonal, 13–8300) for 1 h at 37°C, followed by detection using Alexa Fluor® 555 donkey anti-mouse secondary antibody (1:800, Abcam, 150110) for 1 h at 37°C. The sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, 2 µg/mL, Abcam, ab104139). Images were captured using the Nikon C2 confocal microscope (Nikon, Tokyo, Japan) and analyzed using NIS-Elements software. Antibody specificity was verified (Supplementary Fig. 1, see section on supplementary materials given at the end of this article).

Zhao Y., Namei E., Yang B., Bao X., Sun W., Subudeng G., Cao G., Li H, & Wang G. (2023). Cyclic AMP mediates ovine cumulus–oocyte gap junctional function via balancing connexin 43 expression and phosphorylation. Endocrine Connections, 12(11), e230337.

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Cx43 Immunoprecipitation and Western Blotting

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293T cells were transfected with 2 µg total of the indicated plasmids using JetPRIME reagent (PolyPLUS) according to the manufacturer’s guidelines. Forty-eight hours after transfection, cells were stimulated with 100 ng/mL PMA for 1 hour, the PMA was washed out, and the cells were lysed 5 hours later with a solution containing 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% Triton X-100, 5 mM EDTA, 2 mM sodium orthovanadate, 10 mM iodoacetamide, and protease cocktail inhibitors (Thermo Scientific). After reserving 3% of the cell lysate to run as input, the remainder was pre-cleared with protein A/G agarose beads (Santa Cruz Biotechnology) and then incubated with 1 µg of anti-Cx43 (71–0700, Invitrogen) antibody, 2 µg of anti-HA (H9658, Sigma) antibody, or control anti-rabbit IgG overnight as indicated. The next day, the immunocomplexes were incubated with protein A/G agarose beads (Santa Cruz Biotechnology) for three hours, washed in lysis buffer, eluted with 2X Laemmli sample buffer after boiling for 5 min at 95°C, resolved using SDS-PAGE, and blotted with the indicated antibodies.

Basheer W.A., Harris B.S., Mentrup H.L., Abreha M., Thames E.L., Lea J.B., Swing D.A., Copeland N.G., Jenkins N.A., Price R.L, & Matesic L.E. (2015). Cardiomyocyte-Specific Overexpression of the Ubiquitin Ligase Wwp1 Contributes to Reduction in Connexin 43 and Arrhythmogenesis. Journal of molecular and cellular cardiology, 88, 1-13.

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Immunostaining of Brain Tissue Sections

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4-µm thick sections of formalin-fixed, paraffin-embedded tissue blocks containing either hippocampal regions or the amygdala and basal forebrain were cut and mounted on pre-coated glass slides (Star Frost, Waldemar Knittel GmbH, Braunschweig, Germany). We performed immunostaining for anti-phospho-tau (AT8; mouse monoclonal antibody (MAb); specific for pS202/pT205, 1:200, no pretreatment, Pierce Biotechnology, Rockford, IL, USA), anti-Cx43 (mouse MAb; 1:50, pretreatment pH 6.0 citrate buffer 10 min steamer, Invitrogen, Camarillo, CA, USA), anti-AQP4 (polyclonal rabbit antibody; 1:250, no pretreatment, Sigma-Aldrich, St. Louis, MO, USA), and GFAP (rabbit polyclonal; 1:3,000, pretreatment with Protease 5 min, Dako, Glostrup, Denmark). The antibody reactions were visualized either by DAKO EnVision© (K5007; for Cx4 and AQP4), or EnVision FLEX+© (K8002; for AT8 and GFAP) detection kit. Finally, sections were counterstained with haematoxylin.

Kovacs G.G., Yousef A., Kaindl S., Lee V.M, & Trojanowski J.Q. (2017). Connexin-43 and Aquaporin-4 are markers of ARTAG-related astroglial response. Neuropathology and applied neurobiology, 44(5), 491-505.

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Immunoprecipitation of Connexin 43

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293T cells were transfected with 2 µg total of the indicated plasmids using JetPRIME reagent (PolyPLUS) according to the manufacturer’s guidelines. Forty-eight hours after transfection, cells were stimulated with 100 ng/mL of phorbol 12-myristate 13-acetate (PMA) for 30 min, the PMA was washed out, and the cells were lysed 5.5 hours later with a solution containing 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% Triton X-100, 2.5 mM EDTA, 2 mM sodium orthovanadate, 10 mM iodoacetamide, and protease cocktail inhibitors (Thermo Scientific). After reserving 3.75% of the cell lysate to run as input, the remainder was pre-cleared with protein A/G agarose beads (Santa Cruz Biotechnology) and then incubated with 1 µg of either anti-Cx43 (71–0700, Invitrogen) antibody or control anti-rabbit IgG overnight. The next day, the immunocomplexes were incubated with protein A/G agarose beads (Santa Cruz Biotechnology) for three hours, washed in lysis buffer, eluted with 2X Laemmli sample buffer after boiling for 5 min at 95°C, resolved using SDS-PAGE, and blotted with the indicated antibodies as described above.

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Immunodetection of Cell Adhesion Proteins

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Rabbit polyclonal anti-TMEM43 (N-13) antibody (Santa Cruz, CA, USA) was used in immunofluorescence microscopy and immunoblot analysis to detect TMEM43. We used mouse monoclonal antibodies of anti-α-catenin, anti-ß-catenin, anti-ZO-1, anti-N-cadherin, anti-JUP, and anti-Cx43 (Invitrogen, San Francisco, CA, USA), rabbit anti-Nav1.5 antibody (Abcam, MA, USA) and mouse anti-GAPDH (SIGMA, Saint Louis, Missouri, USA) for detection of these proteins in immunoassays.

Siragam V., Cui X., Masse S., Ackerley C., Aafaqi S., Strandberg L., Tropak M., Fridman M.D., Nanthakumar K., Liu J., Sun Y., Su B., Wang C., Liu X., Yan Y., Mendlowitz A, & Hamilton R.M. (2014). TMEM43 Mutation p.S358L Alters Intercalated Disc Protein Expression and Reduces Conduction Velocity in Arrhythmogenic Right Ventricular Cardiomyopathy. PLoS ONE, 9(10), e109128.

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