DYKDDDDK (binds to Sigma FLAGŪ) (mouse antibody,monoclonal, Clone 1E6)
AS15 3036 | Clonality: Monoclonal | Host: Mouse | Reactivity: DYKDDDDK-tag
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Product Information
Immunogen
KLH-conjugated synthetic peptide: DYKDDDDK (Sigma FLAG®)
Host
Mouse
Clonality
Monoclonal
Purity
Immunoglobulin Protein A/G purified in PBS, pH 7.4. Contains 0.055% sodium azide.
Format
Liquid
Quantity
100 ĩg
Storage
Store at -20°C for up to 1 year, Make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications
Immunoprecipitation (IP), Immunohistochemistry (IHC), Western blot (WB)
Recommended dilution
1 : 100 (WB) 1 : 20 (IP), 1 : 20 (IHC)
Reactivity
Confirmed reactivity
DYKDDDDK epitope tag (Sigma FLAG®), fused to proteins in mammalian cells
Not reactive in
No confirmed exceptions from predicted reactivity are currently known
Application examples
Application examples
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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
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
Additional information
Background
Background
Mouse anti-DYKDDDDK is a primary antibody which binds to DYKDDDDK (Sigma FLAG®).
Product citations
Selected references
Durall et al. (2020). Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803. Biotechnol Biofuels. 2020 Jan 28;13:16. doi: 10.1186/s13068-020-1653-y.
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.
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.
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