Product nameAnti-DDDDK tag antibody - (Equivalent to FLAG antibodies from Sigma)
See all DDDDK tag primary antibodies
DescriptionRabbit polyclonal to DDDDK tag - (Equivalent to FLAG antibodies from Sigma)
Tested applicationsSuitable for: WB, ICC/IF, ELISA, IPmore details
Species reactivityReacts with: Species independent
Peptide xxxDDDDK conjugated to KLH. xxx represents random amino acids. This sequence represents the Enterokinase Cleavage Site.
General notesEpitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with xxxDDDDK may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.2
Preservative: 0.1% Sodium azide
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab1162 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.2 - 1 µg/ml.
Antibody was analyzed by western blot using various amino-terminal and carboxy-terminal DDDDK fusion proteins and Invitrogen Positope" R900-40. A 1/5000 dilution gave bands against 0.1, 1.0 and 10 ng of the fusion proteins and 100 ng and 25 ng of the Positope.
|ICC/IF||Use at an assay dependent concentration.|
|ELISA||Use a concentration of 1 - 5 µg/ml.
As coating antibody in sELISA.
|IP||1/40. PubMed: 25419851|
RelevanceThis is a useful tool for the localisation and characterisation of DDDDK tagged proteins.
- DDDDK epitope tag antibody
- DDDK antibody
- ddk antibody
Immunofluorescent analysis of A293 cells labeling Flag-VPS35 with ab1162 at 10 μg/ml. The nuclear counter stain is DAPI (blue).
ab1162 staining DDDDK tag in hTERT-RPE1 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with Paraformaldehyde, permeabilized with Saponin and blocked with 0.02% BSA for 20 minutes at 18°C. Samples were incubated with primary antibody (1/100) for 45 minutes at 18°C. A Cy3®-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody.
ab1162 staining DDDDK tag (pink) in neonatal rat cardiomyocytes transduced with Ad-TBX18/FLAG-IRES-GFP by Immunocytochemistry/ Immunofluorescence.
Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and incubated with primary antibody. An AlexaFluor®-conjugated anti-rabbit IgG was used as the secondary antibody. Nuclei were stained with DAPI (blue).
This product has been referenced in:
- Li Y et al. Fungal acetylome comparative analysis identifies an essential role of acetylation in human fungal pathogen virulence. Commun Biol 2:154 (2019). Read more (PubMed: 31069264) »
- Frattini V et al. A metabolic function of FGFR3-TACC3 gene fusions in cancer. Nature 553:222-227 (2018). Read more (PubMed: 29323298) »