Tumor tissue was homogenized in radioimmunoprecipitation (RIPA) buffer containing phosphatase inhibitor cocktail (Zmtech scientific). Protein samples (35 µg) were separated on 4–12% Bis-Tris SDS-PAGE gels (Life Technologies), and transferred onto Immuno-Blot PVDF membranes (Bio-Rad). Membranes were blocked in PBS with 5% skim milk and 0.1% Tween-20 for 1 hour, and incubated with primary antibodies overnight at 4°C, then with secondary antibodies for 30 min at room temperature. Protein bands were detected using the ECL Western blotting detection reagent (GE Amersham). Band intensities were quantified using ImageJ and relative protein levels were normalized to β-actin levels. The following antibodies were used: mouse anti-arginase-1 (1:4000, BD Biosciences), mouse anti-β-actin (1:10,000, Sigma), mouse anti-reelin (1:500, Millipore), peroxidase conjugated goat anti-mouse (1:3000, Cell Signaling).
Mouse anti arginase 1
Manufactured by BD
Mouse anti-arginase-1 is a primary antibody that specifically binds to the arginase-1 protein. Arginase-1 is an enzyme involved in the urea cycle. This antibody can be used for the detection and quantification of arginase-1 in various research applications.
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Lab products found in correlation
2 protocols using mouse anti arginase 1
1
Western Blot Analysis of Tissue Proteins
2
Immunoblotting of Cell Lysates
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Total cell lysates were prepared using ice-cold lysis buffer (150 mM NaCl, 50 mM Tris, 1% Nonidet P-40 and 0.1% sodium dodecyl sulfate). The lysates were separated on sodium dodecyl sulfate-polyacrylamide gels and transferred to polyvinylidene difluoride membranes (Bio-Rad, CA). The membranes were blocked with 5% skim milk for 1 h and incubated with mouse anti-arginase-1 (1:1,000 dilution; BD Biosciences, NJ), rabbit anti-microtubule-associated protein light chain 3 (LC3; 1:1,000 dilution; MBL, Woburn, MA), or mouse anti-β actin (1:5,000 dilution; Sigma-Aldrich) antibody overnight at 4°C. Next, the membranes were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies (1:2000 dilution) for 1 h at room temperature. The blots were visualized with a chemiluminescence detection kit (SuperSignal™ West Femto; Thermo Fisher, Franklin, MA).
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