Cells were lysed in Hypotonic Lysis Buffer: 20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM EDTA, 0.5% NP-40, 0.1% Triton X-100, and cOmplete mini protease inhibitor (Sigma-Aldrich) for 20 min on ice. The lysates were mixed 1:1 with 2 × Laemmli buffer containing 1:20-diluted 2-mercaptoethanol, boiled for 10 min, and centrifuged at 16,000 × g for 5 min at 4°C. Samples were run on 4–20% SDS-PAGE and transferred to nitrocellulose membranes. Membrane blocking, as well as antibody binding were in TBS Odyssey Blocking Buffer (Li-Cor, Lincoln, NE). Primary antibodies used were rabbit anti-CypA (1:10,000 dilution; Enzo Life Sciences, Farmingdale, NY, catalogue #BML-SA296) and mouse anti-β-actin (1:1,000 dilution; Abcam, Cambridge, UK, catalogue #ab3280). Goat anti-mouse-680 (Li-Cor, catalogue #925–68070) and goat anti-rabbit-800 (Li-Cor, catalogue #925–32211) as secondary antibodies were used at 1:10,000 dilutions. Blots were scanned on the Li-Cor Odyssey CLx.
Rabbit anti cypa
Manufactured by Enzo Life Sciences
Rabbit anti-CypA is an antibody product used in research applications. It specifically recognizes and binds to the Cyclophilin A (CypA) protein. CypA is a widely expressed protein involved in various cellular processes. The antibody can be utilized in techniques such as Western blotting, immunoprecipitation, and immunohistochemistry to study the expression and localization of CypA.
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Lab products found in correlation
2 protocols using rabbit anti cypa
1
Protein Extraction and Western Blotting
2
Protein Extraction and Western Blotting
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Cells were lysed in Hypotonic Lysis Buffer: 20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM EDTA, 0.5% NP-40, 0.1% Triton X-100, and cOmplete mini protease inhibitor (Sigma-Aldrich) for 20 min on ice. The lysates were mixed 1:1 with 2 × Laemmli buffer containing 1:20-diluted 2-mercaptoethanol, boiled for 10 min, and centrifuged at 16,000 × g for 5 min at 4°C. Samples were run on 4–20% SDS-PAGE and transferred to nitrocellulose membranes. Membrane blocking, as well as antibody binding were in TBS Odyssey Blocking Buffer (Li-Cor, Lincoln, NE). Primary antibodies used were rabbit anti-CypA (1:10,000 dilution; Enzo Life Sciences, Farmingdale, NY, catalogue #BML-SA296) and mouse anti-β-actin (1:1,000 dilution; Abcam, Cambridge, UK, catalogue #ab3280). Goat anti-mouse-680 (Li-Cor, catalogue #925–68070) and goat anti-rabbit-800 (Li-Cor, catalogue #925–32211) as secondary antibodies were used at 1:10,000 dilutions. Blots were scanned on the Li-Cor Odyssey CLx.
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