Der Maus Anti-DYKDDDDK Tag-Antikörper wurde für WB, IP und IF validiert und wurde unabhängig validiert. Er ist geeignet, DYKDDDDK Tag in Proben von zu detektieren.
WB, IP, IF
Wirt: Maus
Monoclonal
F-tag-01
unconjugated
Applikationshinweise
WB (1:1000-10000), IP (1:100-500), IF (1:200-1000). Other applications have not been tested. The optimal dilutions should be determined by end user.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Validierung #028889
(Immunofluorescence)
by
Reveal Biosciences
No.
#028889
Datum
07.11.2013
Antigen
Chargennummer
L21/2013G
Validierte Anwendung
Immunofluorescence
Positivkontrolle
HEK293 cells transfected with DYKDDDDK Tag
Negativkontrolle
Untransfected HEK293 cells
Bewertung
Signal was detected in positive control samples but not in negative control samples.
Validierungsbilder
Figure 1: : Immunofluorescence analysis of cultured cells using DYKDDDDK Tag antibody (Catalog number ABIN1112984, Lot number L21/2013G). Flag is present in the positive control (HEK293 cells transfected with DYKDDDDK Tag panel 1, transfected cells are indicated by green fluorescent staining) and absent from the negative control (untransfected HEK293 cells panel 2). No staining was observed in the isotype (panel 3) and secondary only (panel 4) controls. DAPI was used as a nuclear counterstain (indicated by blue fluorescent staining). Scale bar = 25 µm.
HEK293 cells were grown directly on coverslips and transfected with a DYKDDDDK tagged protein.
Cells were fixed with 4% paraformaldehyde in PBS for 15 min at room temperature (RT).
Fixed cells were rinsed three times in PBS for 5 min each at RT.
Cells were blocked in 1 X PBS / 5% normal goat serum / 0.3% Triton X-100 for 20 min at RT.
Cells were incubated with primary antibody diluted 1:100 in Universal Antibody Dilution Buffer (Electron
Microscopy Sciences, 25885-05) overnight at 4°C.
Cells were rinsed three times in PBS for 5 min each at RT.
Cells were incubated with secondary antibody diluted 1:150 in Universal Antibody Dilution Buffer
(Electron Microscopy Sciences, 25885-05) for 60 min at RT in dark.
Cells were rinsed three times in PBS for 5 min each at RT.
Coverslips were mounted on slides with Fluoro-Gel II with DAPI (Electron Microscopy Sciences Cat. No: 17985-50).
Stained cells were imaged with an EIDAQ 100 fluorescent microscope.
Anmerkungen
None
Format
Liquid
Buffer
PBS (pH7.4), 50% Glycerol and 0.02% Sodium Azide.
Konservierungsmittel
Sodium azide, Thimerosal (Merthiolate)
Lagerung
4 °C
Informationen zur Lagerung
Store at 2-8 °C for 1 month. Or aliquot and store at -20 °C or below for long term. Avoid repeated freeze and thaw cycles.
Target
DYKDDDDK Tag
Andere Bezeichnung
Flag Tag
Substanzklasse
Tag
Hintergrund
DYKDDDDK-Tag, or DYKDDDDK octapeptide, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. The peptide sequence of the DYKDDDDK-Tag is: N-DYKDDDDK-C (1012 Da). It can be used in conjunction with other affinity tags, for example a polyhistidine tag (His-tag), HA-tag or myc-tag. It can be fused to the C-terminus or the N-terminus of a protein.