Isolated plantar arteries opened longitudinally were fixed with 4% paraformaldehyde in PBS (137 mM NaCl, 8.10 mM Na2HPO4·12H2O, 2.68 mM KCl, 147 mM KH2PO4) for 60 min. The specimens were permeabilized with 1.0% Triton X-100 in PBS for 60 min, blocked with 3% BSA fraction V (Roche, Basel, Switzerland) for 30 min, and then incubated with the primary antibody against α-smooth muscle actin (α-SMA; 1:1500, Sigma Aldrich, #A2547) or NCX (1:600, Abcam, #ab151608) in 1% BSA/PBS overnight at 4°C. The specimens were subsequently incubated with goat anti-mouse IgG antibody-conjugated Alexa Fluor 546 (1:1000, Invitrogen, Carlsbad, CA, USA, #A11003), goat anti-rabbit IgG antibody-conjugated Alexa Fluor 488 (1:1000, Invitrogen, #A11034), and hoechst 33342 (1:3000, Dojindo, Kumamoto, Japan) in 1% BSA/PBS for 60 min at room temperature. Fluorescence microscopy images were obtained on a Nikon A1 RSi (Nikon, Minato City, Tokyo, Japan).
Ab151608
Manufactured by Abcam
Sourced in Switzerland
Ab151608 is a recombinant monoclonal antibody that recognizes human CD8 alpha. The antibody is designed for use in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ab151608
1
Immunofluorescence Staining of Plantar Arteries
2
Analyzing Cardiac Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein lysates from mouse heart tissues were harvested in radioimmunoprecipitation assay buffer (RIPA, 50 mM Tris-HCl; pH7.4, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 150 mM NaCl, 2 mM EDTA), supplemented with protease and phosphatase inhibitors (Roche), for 30 mins on ice, spun down at 15,000 × g at 4 ° C. Soluble fractions were saved for immunoblotting. Protein lysates were heated for 5 min to either 65 °C or to boiling, run on 4–12% polyacrylamide gels and transferred onto 0.22–0.45 μm nitrocellulose membranes. After blocking for 1 hour with 5% milk in 0.05% TBS-Tween20, primary antibodies were added and incubated at 4 ° C overnight: primary mouse monoclonal anti-DHPR (1:500 dilution; ab2864; Abcam), primary mouse monoclonal anti-RyR2 (1:1000 dilution; ab2827; Abcam), primary mouse monoclonal anti-PLN (1:1000 dilution; MA3-922; ThermoFisher), primary mouse monoclonal anti-SERCA2A (1:1000 dilution; MA3-919; ThermoFisher), and primary rabbit polyclonal anti-NCX1 (1:1000 dilution; ab151608; Abcam).
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