DYKDDDDK Tag antibody

Details for Product No. ABIN1112984, Supplier: Log in to see
Antigen
Reactivity
Tag
240
101
39
18
18
9
8
7
4
3
2
1
1
1
Host
Mouse
173
110
34
34
17
Conjugate
Un-conjugated
38
21
16
8
8
8
8
6
6
5
5
5
5
5
5
5
4
3
3
3
2
2
2
1
1
1
1
1
1
1
1
1
Application
Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
289
131
126
103
84
58
43
14
11
8
7
7
4
4
4
4
3
3
3
2
1
1
1
1
1
1
Options
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Available images

'Independent Validation' Badge
Lot Number L21/2013G
Method validated Immunofluorescence
Positive Control HEK293 cells transfected with DYKDDDDK Tag
Negative Control Untransfected HEK293 cells
Notes Signal was detected in positive control samples but not in negative control samples.
Primary Antibody
  • Antibody: DYKDDDDK Tag antibody
  • Catalog number: ABIN1112984
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: L21/2013G
Secondary Antibody
  • Antibody: Goat anti-mouse IgG (H+L) IF488 conjugated antibody
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: S6/2013S
Controls
  • 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.
Protocol
  • 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.
Experimental Notes None
Validation Images
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.
Sequence DYKDDDDK
Isotype IgG1
Alternative Name Flag Tag
Target Type Tag
Background 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.
Application Notes 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.
Plate Strips (12 x 8)
Restrictions For Research Use only
Format Liquid
Buffer PBS (pH7.4), 50% Glycerol and 0.02% Sodium Azide.
Preservative Sodium azide, Thimerosal (Merthiolate)
Storage 4 °C
Storage Comment 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.