Anti‐GFP‐HRP and anti‐FLAG‐HRP antibodies were obtained from Miltenyi Biotec, USA. Ubiquitinated H2B monoclonal antibody (NRO3) was obtained from Medimabs, Canada. Western blots were performed using the Bio‐Rad trans‐blot turbo system using polyvinylidene difluoride (PVDF) membranes. The blots were blocked using TBS/2% BSA/0.3% Tween‐20 and 1 h shaking at room temperature. Here, 1 : 5000 dilutions of the antibodies were used for detection using chemiluminescence and the Clarity Western ECL substrate (Bio‐Rad). For detection of H2Bub, a secondary anti‐rabbit‐HRP antibody was used (1 : 5000 dilution).
Anti flag hrp antibodies
Manufactured by Miltenyi Biotec
Sourced in United States
Anti-FLAG-HRP antibodies are detection reagents used to identify and visualize proteins tagged with the FLAG epitope. The antibodies are conjugated with horseradish peroxidase (HRP), which allows for sensitive chemiluminescent or colorimetric detection of FLAG-tagged proteins.
Automatically generated - may contain errors
2 protocols using anti flag hrp antibodies
1
Western Blot Analysis of Ubiquitinated H2B
2
Protein Interaction Identification Using Co-IP
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FLAG-tagged SPX1 and SPX3 and GFP-tagged PHR2 (Medtr1g080330) constructs were transiently expressed in Nicotiana leaves as described (Zeng et al., 2018) . Total proteins were isolated using Co-IP buffer (10% glycerol, 50 mM Tris-Hcl pH=8.0, 150 mM NaCl, 1%
Igepal CA 630, 1 mM PMSF, 20 µM MG132, 1 tablet protease inhibitor cocktail). GFP-Trap agarose beads (Chromotek) were used to immunoprecipitate GFP protein complexes. Western blot was performed as described (Bungard et al., 2010) . 1:5000 diluted anti-GFP-HRP and anti-FLAG-HRP antibodies (Miltenyi biotec, USA) were used for detection.
Igepal CA 630, 1 mM PMSF, 20 µM MG132, 1 tablet protease inhibitor cocktail). GFP-Trap agarose beads (Chromotek) were used to immunoprecipitate GFP protein complexes. Western blot was performed as described (Bungard et al., 2010) . 1:5000 diluted anti-GFP-HRP and anti-FLAG-HRP antibodies (Miltenyi biotec, USA) were used for detection.
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