Infected cells (~1.5–2 × 106 cells/mL, MOI = 3) were collected at 0, 24, 48, and 72 hpi by centrifugation at 1000× g for 10 min at 4 °C. The cells were lysed in RIPA buffer (Fisher Scientific), quantified by BCA assay (Fisher Scientific), and ~10 µg of protein was separated by electrophoresis in 10% TGX Stain-Free precast mini SDS-PAGE gels (Bio-Rad, Mississauga, ON, Canada) according to manufacturer’s directions. After transfer to PVDF membranes, Western blot analysis was performed with anti-Cas9 (MAC133; Sigma-Aldrich) or anti-GP64 (AcV5, Fisher Scientific) as primary antibodies and goat anti-mouse IgG HRP secondary (Bio-Rad) and imaged on a ChemiDoc MP Imager (Bio-Rad). The Image Lab software was used for further image processing (Bio-Rad).
Anti gp64 acv5
Manufactured by Thermo Fisher Scientific
Anti-GP64 (AcV5) is a monoclonal antibody that recognizes the baculovirus envelope glycoprotein GP64. It can be used for the detection and analysis of baculovirus-infected insect cells.
Automatically generated - may contain errors
Lab products found in correlation
Image lab software, by Bio-Rad (3 mentions)
Ripa buffer, by Thermo Fisher Scientific (3 mentions)
Chemidoc mp imager, by Bio-Rad (3 mentions)
Goat anti mouse igg hrp secondary, by Bio-Rad (3 mentions)
Bca assay, by Thermo Fisher Scientific (2 mentions)
Tgx stain free precast mini sds page gels, by Bio-Rad (2 mentions)
Anti cas9 mac133, by Merck Group (2 mentions)
Pierce bca assay, by Thermo Fisher Scientific (1 mentions)
3 protocols using anti gp64 acv5
1
Quantifying Cas9 and GP64 Protein Levels
2
Quantifying Baculovirus Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Infected cells ( 1.5–2 cells/mL) were collected at 20–24 hpi by centrifugation at 500 for 10 min at 4 . The cells were lysed in RIPA buffer (Fisher Scientific), quantified by Pierce BCA assay (Fisher Scientific), and 10 g of protein was separated by electrophoresis in 10% TGX Stain-Free precast mini SDS-PAGE gels (Bio-Rad, Mississauga, ON, Canada) according to manufacturer’s directions. After transfer to low fluorescence PVDF membranes, Western blot analysis was performed with anti-GP64 (AcV5, Fisher Scientific) primary antibody and goat anti-mouse IgG HRP secondary (Bio-Rad) and imaged on a ChemiDoc MP Imager (Bio-Rad). The Image Lab software (Bio-Rad) was used for further image processing.
3
Cas9 and GP64 Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Infected cells (˜1.5-2x10 6 cells/ml, MOI = 3) were collected at 0, 24, 48, and 72 hpi by centrifugation at 1000 x g for 10 min at 4 degC. The cells were lysed in RIPA buffer (Fisher Scientific), quantified by BCA assay (Fisher Scientific), and ˜10 μg of protein was separated by electrophoresis in 10\% TGX Stain-Free precast mini SDS-PAGE gels (Bio-Rad, Mississauga ON) according to manufacturer's directions. After transfer to PVDF membranes, Western blot analysis was performed with anti-Cas9 (MAC133; Sigma-Aldrich) or anti-GP64 (AcV5, Fisher Scientific) as primary antibodies and goat anti-mouse IgG HRP secondary (Bio-Rad) and imaged on a ChemiDoc MP Imager (Bio-Rad). The Image Lab software was used for further image processing (Bio-Rad).
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