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

DYKDDDDK (binds to Sigma FLAGŪ) in the group Tag Antibodies / DYKDDDDK (binds to SigmaFLAGŪ) at Agrisera AB (Antibodies for research) (AS15 2871)


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Info: Add review
product information
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.


KLH-conjugated DYKDDDDK (FLAG) synthetic peptide

Host Mouse
Clonality Monoclonal
Clone IgG1, clone FG4R
Purity Protein A purified
Format Lyophilized in 10mM PBS, pH 7.2, with 1% BSA and 0.05% sodium azide
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl of sterile water.

Lyophilized antibody can be stored at -20°C for up to 3 years. Re-constituted antibody can be stored at 4°C for several days to weeks.

Once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Dot blot (Dot), ELISA (ELISA), Immunohistochemistry (IHC), Immunoprecipitation (IP), Western blot (WB)
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other tag antibodies

Plant and algal protein extraction buffer

Secondary antibodies

Additional information Working dilution for Dot Blot, ELISA and IP needs to be determined experimentally.
application information
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
Predicted reactivity
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information Antibody is purified on Protein A
Selected references

to be added when available antibody available in July 2015

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

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