DYKDDDDK (binds to Sigma FLAGŪ)
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AS15 2871 | Clonality: monoclonal | Host: Mouse | Reactivity: N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
|Recommended dilution||1 : 1000-5000 (1-5ng) (WB) (incubate for one hour at room temperature). 1 : 500-1 :2000 (IS) For best results with other assays (Dot, ELISA, IP), please determine optimal working dilution by end user|
|Expected | apparent MW|
|Confirmed reactivity||N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Additional information||Antibody is purified on Protein A|
to be added when available antibody available in July 2015
20 µg of total protein from Chlamydomonas reinhardtii Flag; 2 (1) 1xFlag 5’+ICL (1x FLAG coding sequence : GA-CTA-CAA-GGA-CGA-CGA-TGA-CAA-G) (2); 1xFlag 5’+ICL (1x FLAG codingsequence :GA-CTA-CAA-GGA-CGA-CGA-TGA-CAA-G) (3); ICL+ 3xFlag 3’end (3xFLAG codingsequence : GAC TAC AAG GAC CAC GAC GGT GAC TAC AAG GAC CAC GAC ATC GAC TAC AAG GAC GACGACGAC AAG TGA.) (4); 1xFlag 5’+ICL+ 3xFlag 3’end (5) were separated on 12 % SDS-PAGE and blotted ON (30V) to PVDF using tank transfer. Blots were blocked with 5 % non-fat milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-mouse IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with BM ECL kit according to the manufacturer's instructions. Exposure time was seconds.
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