2 million macrophages were lysed in 200 μL of RIPA buffer (Tris-HCl, pH 7.4, 50 mM, NaCl 150mM, NP-40 1%, sodium deoxycholate 0.5%, SDS 0.1%) with a cocktail of protease and phosphatase inhibitors (Roche). 20 μg of protein was loaded per lane and separated on a 4%–12% SDS/PAGE gel and transferred to PVDF. Ponceau stain (Sigma) determined protein transfer and membrane was blocked for 1 h with 5% skim milk in TBS/0.1% Tween at 25°C, and then incubated with the indicated primary antibody in 3% BSA TBS/0.1% Tween overnight at 4°C. Specific antibodies Influenza NP protein (A. Sastre-Garcia) were used to quantify virus protein in the lung tissue. β-actin (Sigma) was used as a loading control. Anti-pan-AGO for immunoblotting (7G1.1) was from the Darnell laboratory (Rockefeller University), anti-AGO1 antibody was from Abcam.
Anti pan ago for immunoblotting 7g1.1
Manufactured by Abcam
Anti-pan-AGO for immunoblotting (7G1.1) is a primary antibody designed for the detection of Argonaute (AGO) proteins in immunoblotting applications. The antibody recognizes a conserved epitope shared among all four AGO proteins, enabling the simultaneous detection of multiple AGO isoforms.
Automatically generated - may contain errors
2 protocols using anti pan ago for immunoblotting 7g1.1
1
Quantifying Influenza Viral Proteins in Macrophages
2
Quantifying Influenza Viral Proteins in Macrophages
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2 million macrophages were lysed in 200 μL of RIPA buffer (Tris-HCl, pH 7.4, 50 mM, NaCl 150mM, NP-40 1%, sodium deoxycholate 0.5%, SDS 0.1%) with a cocktail of protease and phosphatase inhibitors (Roche). 20 μg of protein was loaded per lane and separated on a 4%–12% SDS/PAGE gel and transferred to PVDF. Ponceau stain (Sigma) determined protein transfer and membrane was blocked for 1 h with 5% skim milk in TBS/0.1% Tween at 25°C, and then incubated with the indicated primary antibody in 3% BSA TBS/0.1% Tween overnight at 4°C. Specific antibodies Influenza NP protein (A. Sastre-Garcia) were used to quantify virus protein in the lung tissue. β-actin (Sigma) was used as a loading control. Anti-pan-AGO for immunoblotting (7G1.1) was from the Darnell laboratory (Rockefeller University), anti-AGO1 antibody was from Abcam.
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