SDS-PAGE and protein blotting were performed as described previously (van Dooren et al., 2008 (link)), except that membranes used for neutravidin blotting were blocked with 3% (w/v) bovine serum albumin (BSA). Blue native PAGE was performed using the NativePAGE system (Thermo Scientific) as described previously (van Dooren et al., 2016 (link)). Blots were probed with antibodies against mouse anti-c-myc (1:500 dilution; Santa Cruz clone 9E10), rabbit anti-Hsp60 (1:1000 dilution; Tonkin et al., 2004 (link)), rabbit anti-T. gondii cytochrome c (1:500 dilution; E.T. and G.v.D., unpublished), rat anti-HA (1:500 to 1:1000 dilution; Sigma clone 3F10), mouse anti-FLAG (1:500 to 1:2000 dilution; Sigma clone M2), and rabbit anti-AtpB (1:500; Agrisera, catalog number AS05 085). Horseradish peroxidase (HRP)-conjugated anti-mouse, anti-rat and anti-rabbit antibodies (Santa Cruz) were used at 1:5000 dilution. For probing for mouse antibodies on immunoprecipitation western blots, HRP-conjugated anti-mouse TrueBlot ULTRA antibodies (eBioscience) were used at 1:5000 dilution. Neutravidin-HRP (Life Technologies) was used to detect biotinylated proteins on membranes at 1:10,000 dilution.
Hrp conjugated anti mouse trueblot ultra antibodies
Manufactured by Thermo Fisher Scientific
HRP-conjugated anti-mouse TrueBlot ULTRA antibodies are laboratory reagents designed for immunodetection applications. They contain horseradish peroxidase (HRP) conjugated to antibodies that specifically recognize mouse immunoglobulins.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti flag, by Merck Group (2 mentions)
Nativepage system, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated goat anti mouse igg, by Abcam (1 mentions)
Goat anti rat igg, by Abcam (1 mentions)
Rat anti ha, by Merck Group (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Goat anti rabbit igg, by Abcam (1 mentions)
2 protocols using hrp conjugated anti mouse trueblot ultra antibodies
1
Protein Analysis by Electrophoresis and Blotting
2
Protein complex analysis by electrophoresis
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Sodium dodecylsulfate (SDS)-polyacrylamide electrophoresis (PAGE), blue native (BN)-PAGE and immunoblotting were performed as described previously [18, (link)41] (link). Primary antibodies used included mouse anti-FLAG (1:100-1:2,000 dilution; Sigma clone M2), rat anti-HA (1:2000 dilution; Sigma clone 3F10), and rabbit anti-Tom40 (1:2,000 dilution; [18] (link)). The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Abcam), goat anti-rabbit IgG (Abcam) and goat anti-rat IgG (Abcam). For probing for mouse antibodies on immunoprecipitation western blots, HRP-conjugated anti-mouse TrueBlot ULTRA antibodies (eBioscience) were used at 1:2,500 dilution. Blots were imaged using X-ray film or using a ChemiDoc MP imaging system (BioRad).
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