Three confluent 160-mm-diameter dishes of MBECs were harvested and plasma membranes were isolated as previously described (Yao et al., 2009). Protein concentrations were determined by Bio-Rad Dc-Protein assay (Bio-Rad). Samples were mixed with Bio-Rad XT sample buffer and reducing agent, run on Criterion™ XT 4–12% Bis-Tris Gels (Bio-Rad), and transferred onto PVDF membranes. Immunodetection was performed with (1:100) rabbit anti-connexin 43 antibody (Invitrogen), (1:500) mouse anti phospho-caveolin1 (pY14) (BD Transduction Laboratories), (1:5000) rabbit anti-β-catenin (Abcam), (1:1000) rabbit anti-Cav-1 (Abcam), or (1:1000) mouse-anti-transferrin receptor (Life Technology) followed by HRP-coupled secondary antibodies (Jackson ImmunoResearch), and developed with ECL-Plus (GE Healthcare). Protein bands were quantified by densitometry using Image Lab 5.1 (Bio-Rad) software. Plasma membrane connexin43 levels were normalized to plasma membrane transferrin receptor. This value was normalized to total β-catenin as a housekeeping reference protein.
Rabbit anti connexin 43 antibody
Manufactured by Thermo Fisher Scientific
The Rabbit anti-connexin 43 antibody is a laboratory tool used to detect and study the protein connexin 43 in various research applications. Connexin 43 is a type of gap junction protein that plays a role in cell-to-cell communication. This antibody can be used to identify and analyze the expression and distribution of connexin 43 in different cell types and tissues.
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Lab products found in correlation
Hrp coupled secondary antibody, by Jackson ImmunoResearch (2 mentions)
Image lab 5, by Bio-Rad (2 mentions)
Rabbit anti cav 1, by Abcam (2 mentions)
Mouse anti transferrin receptor, by Thermo Fisher Scientific (2 mentions)
Criterion xt 4 12 bis tris gel, by Bio-Rad (2 mentions)
Mouse anti phospho caveolin1 py14, by BD (2 mentions)
Dc protein assay, by Bio-Rad (2 mentions)
Rabbit anti β catenin, by Abcam (2 mentions)
Ecl plus, by GE Healthcare (2 mentions)
2 protocols using rabbit anti connexin 43 antibody
1
Plasma Membrane Protein Analysis
2
Plasma Membrane Protein Analysis
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Three confluent 160-mm-diameter dishes of MBECs were harvested and plasma membranes were isolated as previously described (Yao et al., 2009). Protein concentrations were determined by Bio-Rad Dc-Protein assay (Bio-Rad). Samples were mixed with Bio-Rad XT sample buffer and reducing agent, run on Criterion™ XT 4–12% Bis-Tris Gels (Bio-Rad), and transferred onto PVDF membranes. Immunodetection was performed with (1:100) rabbit anti-connexin 43 antibody (Invitrogen), (1:500) mouse anti phospho-caveolin1 (pY14) (BD Transduction Laboratories), (1:5000) rabbit anti-β-catenin (Abcam), (1:1000) rabbit anti-Cav-1 (Abcam), or (1:1000) mouse-anti-transferrin receptor (Life Technology) followed by HRP-coupled secondary antibodies (Jackson ImmunoResearch), and developed with ECL-Plus (GE Healthcare). Protein bands were quantified by densitometry using Image Lab 5.1 (Bio-Rad) software. Plasma membrane connexin43 levels were normalized to plasma membrane transferrin receptor. This value was normalized to total β-catenin as a housekeeping reference protein.
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