DYKDDDDK (binds to Sigma FLAGŪ) (mouse antibody,monoclonal, Clone 1E6)
AS15 3036 | Clonality: Monoclonal | Host: Mouse | Reactivity: DYKDDDDK-tag
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KLH-conjugated synthetic peptide: DYKDDDDK (Sigma FLAG®)
1 and 2. pBA002 vector as negative control (technical replicates).
3 and 4. pBA002:AtACO4 (technical replicates)
18 µg/well of total protein extracted freshly from Nicotiana benthamiana with following buffer components: Tris-HCl pH 7.5 100 mM, Urea 8 M, Triton x-100 (0.2%), Sarkosyl (0.2%), PMSF 1mM, Pepstatin 1 µg/mL, Leupeptin (1 µg/mL), NEM 2 mM and Iodoacetamida 10 mM and denatured with 4x Laemmli Sample Buffer at 95 °C for 5 min. Protein samples were separated on 12% SDS-PAGE and blotted for 1h to PVDF (pore size of 0.2 µm), using semi-dry transfer. Blot was blocked with 3% milk in TBS-T for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 (anti-ACO4) or 1:2000 (anti-flag, Agrisera, No. AS15 3036, Lot. 1902) in 3% milk in TBS-T ON/4°C with agitation (anti-ACO4 and anti-FLAG in corresponding membrane). The antibody solution was removed and the blot was washed three times for 10 min in TBS-T at RT with agitation. Blot was incubated with Peroxidase labelled anti-mouse antibody diluted to 1:25 000 for 1h/RT with agitation. The blot was washed as above and developed for 30 min with AgriseraECLSuperBright. Exposure time was 60 seconds. Asterisk denotes a band recognized in all samples.
Courtesy of Dr. Maria Lois, CRAG, Spain
Mouse anti-DYKDDDDK is a primary antibody which binds to DYKDDDDK (Sigma FLAG®).
Wardhan et al. (2017). Chickpea transcription factor CaTLP1 interacts with protein kinases, modulates ROS accumulation and promotes ABA-mediated stomatal closure. Sci Rep. 2016 Dec 9;6:38121. doi: 10.1038/srep38121.
Related products: DYKDDDDK (binds to Sigma FLAGŪ) (mouse antibody,monoclonal, Clone 1E6)
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