DYKDDDDK (binds to Sigma FLAG®, clone FG4R)

Product no: AS15 2871

AS15 2871  | Clonality: monoclonal  |  Host: Mouse  |  Reactivity: N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins

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  • Product Info
  • Sub class: IgG1

    KLH-conjugated DYKDDDDK (FLAG) synthetic peptide

    Host: Mouse
    Clonality: Monoclonal
    Purity: Total IgG. Protein A purified in 10mM PBS, pH 7.2, with 1% BSA and 0.05% sodium azide.
    Format: Liquid
    Quantity: 50 µg
    Storage: Store at -20°C for up to 2 years.
    Tested applications: Dot blot (Dot), ELISA (ELISA), Immunohistochemistry (IHC), Immunoprecipitation (IP), Western blot (WB)
    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
  • Reactivity
  • Confirmed reactivity: N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • application example

    western blot using anti-FLAG antibody

    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.

    Courtesy of Dr. Nadine Coosemans, Remacle lab, University of Liège, Belgium
  • Additional Information
  • Additional information: Working dilution for Dot Blot, ELISA and IP needs to be determined experimentally.

    Antibody is purified on Protein A.
  • Background
  • Background: FLAG-tag is a tag that can be added to a protein sequence motif DYKXXD. It can be used for affinity chromatography, to separate recombinant protein from wt proteins. It can also be used to isolate protein complexes with multiple subunits.

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