DYKDDDDK Tag Antikörper

Details zu Produkt Nr. ABIN1112984, Anbieter: Anmelden zum Anzeigen
Antigen
Reaktivität
Tag
238
101
65
18
18
9
8
7
4
3
2
1
1
1
Wirt
Maus
198
109
34
34
17
Konjugat
Unkonjugiert
39
21
18
10
10
10
8
8
7
7
7
7
7
7
6
6
5
3
3
3
2
2
2
1
1
1
1
1
1
1
1
1
Applikation
Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
313
131
125
103
84
58
43
14
11
8
7
7
4
4
4
4
3
3
3
2
1
1
1
1
1
1
Optionen
Hersteller
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'Independent Validation' Siegel
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.
Primärantikörper
Sekundärantikörper
Kontrollen
  • Positive control: HEK293 cells from ATCC transfected with a DYKDDDDK tagged protein (specimen
  • known to contain the target protein).
  • Negative control: Untransfected HEK293 cells from ATCC (specimen known to not contain the target
  • protein).
  • Isotype control: HEK293 cells from ATCC transfected with a DYKDDDDK tagged protein treated with
  • isotype mouse IgG control instead of the primary antibody. Any staining observed is due to non-specific
  • binding of secondary antibody.
  • Secondary antibody only control: HEK293 cells from ATCC transfected with a DYKDDDDK tagged
  • protein treated with Goat anti-Mouse IF488 secondary antibody only. Any staining observed is due to
  • non-specific binding of secondary antibody.
Protokoll
  • 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
Validierungsbilder
Immunofluorescence validation image for anti-DYKDDDDK Tag antibody (ABIN1112984) Figure 1: : Immunofluorescence analysis of cultured cells using DYKDDDDK Tag antibody...
Immunogen A synthetic peptide DYKDDDDK conjugated to KLH.
Sequenz DYKDDDDK
Isotyp IgG1
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.
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.
Plattentyp Strips (12 x 8)
Beschränkungen Nur für Forschungszwecke einsetzbar
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.
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