Cell lysates were boiled in 1× Laemmli sample buffer for 5 min, and then electrophoresed on 4–20% Tris-Glycine eXtended (TGX) precast protein gels (Biorad). Proteins were transferred to 0.2 μm polyvinylidene difluoride (PVDF) membranes by using a Trans-Blot Turbo Transfer System (Biorad). The membranes were blocked for 30 min in 5% non-fat milk, and then incubated for 2 hours at room temperature or overnight at 4 °C with a primary antibody. After 3 times of 5-min wash with TBST, the membranes were immunoblotted with alkaline phosphatase (AP) conjugated goat-anti-rabbit IgG or goat-anti-mouse IgG secondary antibody (Promega) for 1 hour at room temperature. After 3 times of 5-min wash with TBST, blots were developed using 5-Bromo-4-chloro-3-indolyl phosphate (BCIP, ThermoFisher) and Nitro blue tetrazolium (NBT, ThermoFisher) as substrates. The blots were then imaged on a ChemiDoc Imaging System (Biorad) and the bands on blots were quantified by ImageJ (Open source Java program from NIH) or the immunoblotting quantification software Image Studio™ Lite (LI-COR Biosciences).
Ap conjugated goat anti rabbit igg or goat anti mouse igg secondary antibody
Manufactured by Promega
The AP-conjugated goat-anti-rabbit IgG or goat-anti-mouse IgG secondary antibody is a reagent used in immunoassay and immunohistochemistry techniques. It is designed to detect and bind to primary antibodies raised in rabbit or mouse hosts, enabling the visualization and quantification of target proteins or antigens.
Automatically generated - may contain errors
2 protocols using ap conjugated goat anti rabbit igg or goat anti mouse igg secondary antibody
1
Western Blot Analysis Protocol
2
Western Blot Analysis Protocol
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Cell lysates were boiled in 1× Laemmli sample buffer for 5 min, and then electrophoresed on 4–20% Tris-Glycine eXtended (TGX) precast protein gels (Biorad). Proteins were transferred to 0.2 μm polyvinylidene difluoride (PVDF) membranes by using a Trans-Blot Turbo Transfer System (Biorad). The membranes were blocked for 30 min in 5% non-fat milk, and then incubated for 2 hours at room temperature or overnight at 4 °C with a primary antibody. After 3 times of 5-min wash with TBST, the membranes were immunoblotted with alkaline phosphatase (AP) conjugated goat-anti-rabbit IgG or goat-anti-mouse IgG secondary antibody (Promega) for 1 hour at room temperature. After 3 times of 5-min wash with TBST, blots were developed using 5-Bromo-4-chloro-3-indolyl phosphate (BCIP, ThermoFisher) and Nitro blue tetrazolium (NBT, ThermoFisher) as substrates. The blots were then imaged on a ChemiDoc Imaging System (Biorad) and the bands on blots were quantified by ImageJ (Open source Java program from NIH) or the immunoblotting quantification software Image Studio™ Lite (LI-COR Biosciences).
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