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DYKDDDDK (binds to Sigma FLAG®) (polyclonal)
AS20 4442 | Clonality: Polyclonal | Host: Rabbit | Reactivity: DYKDDDDK-Tag
- Product Info
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Immunogen: KLH-conjugated synthetic peptide: DYKDDDDK (Sigma FLAG®) Host: Rabbit Clonality: Polyclonal Purity: Immunogen affinity purified serum, in PBS pH 7.4. Format: Lyophilized Quantity: 50 µg Reconstitution: For reconstitution add 50 µl, of sterile water. Storage: Store lyophilized/reconstituted at -20°C; once reconstituted 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: Western blot (WB) Recommended dilution: 1 : 1000 - 1: 5000 (WB) Expected | apparent MW: Depends upon fusion protein partner - Reactivity
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Confirmed reactivity: DYKDDDDK epitope tag (Sigma FLAG®), fused to proteins in plant cells Not reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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Recombinant FLAG-protein was expressed in bacteria and purified with affinity chromatography. 1.5 µg of FLAG protein was used per in vitro SUMOylation assay in two different conditions: line A. in 30°C for 3 hours and in B: 30°C overnight. FLAG-protein-SUMO1 FLAG-protein Samples were separated on 10% SDS-PAGE and transferred to PVDP membrane by semidry blotting. Membrane was blocked with 5% skim milk in PBST (0.01% Tween 20) for 1h in RT with agitation. Then blot was incubated with primary antibody anti FLAG at dilution of 1:5000 in 5% skim milk/PBST for 1h in RT with agitation. After washes (3 times for 5 min. in PBST) blot was incubated with secondary antibody (anti rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) at dilution 1:25 000 for 1h in RT also with 5% skim milk. The blot was washed 3 times for 5 min. with PBST and developed for 2 min with ECL according to the manufacturer's instructions. Exposure time was 1.5 min.
Dr Agata Cieśla, Ludwików's lab, Laboratory of Biotechnology UAM, Poland - Background
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Background: Rabbit anti-DYKDDDDK is a primary antibody which binds to DYKDDDDK (Sigma FLAG®). - Protocols
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts
Agrisera Educational Poster Collection - Reviews:
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