DDDDK-Tag Rabbit Monoclonal Antibody
| Cat Number: | MAB-94792 |
|---|---|
| Conjugate: | Unconjugated |
| Size: | 100 ug |
| Clone: | ARC5111-01 |
| Host: | Rabbit |
| Isotype: | IgG |
| Immunogen: | A synthetic peptide corresponding to DDDDK tag. |
| Reactivity: | Species independent |
| Applications: | Western Blot: 1:2000 – 1:10000 |
| Molecular Weight: | 56kDa/50kDa/46kDa/68kDa |
| Purification: | Affinity purification |
| Synonyms: | DDDDK;DDDDK tag;DDDDK-tag |
| Background: | FLAG-tag, or FLAG octapeptide, or FLAG epitope, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology, having the sequence motif DYKDDDDK. It has been used for studying proteins in living cells and for protein purification by affinity chromatography. It has been used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits, because its mild purification procedure tends not to disrupt such complexes. It has been used to obtain proteins of sufficient purity and quality to carry out 3D structure determination by x-ray crystallography.A FLAG-tag can be used in many different assays that require recognition by an antibody. If there is no antibody against a given protein, adding a FLAG-tag to a protein allows the protein to be studied with an antibody against the FLAG sequence. Examples are cellular localization studies by immunofluorescence or detection by SDS PAGE protein electrophoresis and Western blotting. |
| Form: | Liquid |
| Buffer: | PBS with 0.05% proclin300,0.05% BSA,50% glycerol,pH7.3. |
| Storage: | Store at -20℃. Avoid freeze / thaw cycles. |
Western blot analysis of 293T, 293T transfected with GSK3B Protein , 293T transfected with BCAT2-N Protein and 293T transfected with BCAT2-C Protein, using DDDDK-Tag antibody at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL West Pico Plus.
Exposure time: 10s.
Western blot analysis of 293F, 293F transfected with COPB2-N Protein and 293F transfected with COPB2-C Protein, using DDDDK-Tag antibody at 1:10000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL West Pico Plus.
Exposure time: 30s.
Immunoprecipitation analysis of 300ug extract cell lysate from 293T cells transfected with GSK3B expression vector containing a DDDDK-Tag with 3 μg DDDDK-Tag Rabbit mAb antibody .Western blot was performed from the immunoprecipitate using DDDDK-Tag Rabbit mAb antibody at a dilution of 1:10000.
Immunofluorescence analysis of 293TFlag- C and 293T-Flag-N and 293T cells using DDDDK-Tag Rabbit mAb at dilution of 1:100 (40x lens).
Blue: DAPI for nuclear staining.
Chromatin immunoprecipitation analysis of extracts of 293T cells transfected with GATA3-DDDDKTag,
using DDDDK-Tag Rabbit mAb antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.
Flow cytometry: 1X10^6 293T cells (negative control,left) and 293T (Transfection,right) cells were surfacestained with DDDDK-Tag Rabbit mAb 2.5 μg/mL orange line or Alexa Fluor 488 Rabbit IgG isotype control 2.5μg/mL, blue line.
Non-fluorescently stained cells were used as blank control (red line).