IHC
Immunohistochemical analysis of paraffin-embedded HeLa transfected with FLAG-tagged Claudin 18.2 (C-terminal) cells with Rabbit anti-DYKDDDDK Tag (FLAG) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling DYKDDDDK Tag (FLAG) with Rabbit anti-DYKDDDDK Tag (FLAG) antibody at 1/10,000 dilution. HeLa cells, transfected with FLAG-tagged empty control, Claudin 18.2 (C-terminal) or Histone H3.1 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-DYKDDDDK Tag (FLAG) antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HeLa cells transfected with FLAG-tagged Otx1 (C-terminal) labeling DYKDDDDK Tag (FLAG). Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
DYKDDDDK Tag (FLAG) was immunoprecipitated in 2µg L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate with Rabbit anti-DYKDDDDK Tag (FLAG) antibody. Western blot was performed from the immunoprecipitate using DYKDDDDK Tag (FLAG) at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/100,000 dilution was used for 1 hour at room temperature. Lane 1: L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate (input). Lane 2: Rabbit IgG instead of Rabbit anti-DYKDDDDK Tag (FLAG) antibody IP in L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate. Lane 3: Rabbit anti-DYKDDDDK Tag (FLAG) antibody IP in L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 17 seconds| Product Name | DYKDDDDK Tag (FLAG) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIP |
| IHC:1:1000 ICC:1:10000 FC:1:10000 IP:1-2ug/sample |
|
| Species Reactivity | A wide range of other species |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | Flag-tag |
|---|---|
| Gene Synonyms | Flag tag Flag-tag DDDDK TAG DDDDK-TAG DYKDDDDK tag DYKDDDDK-tag |
| Gene Full Name | FLAG-tag |
| Gene Summary | The DYKDDDDK peptide (Flag-tag) is a polypeptide protein tag that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, and then used to separate recombinant, over expressed protein from wild-type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits. |
| Alternative Names | DDDDK epitope tag antibody DDDK antibody ddk antibody DYKDDDDK antibody DYKDDDDK epitope tag antibody DYKDDDDK tag antibody ECS epitope tag antibody ECS tag antibody Enterokinase Cleavage Site epitope tag antibody Enterokinase Cleavage Site tag antibody DDDDK epitope tag antibody DDDK antibody ddk antibody DYKDDDDK antibody DYKDDDDK epitope tag antibody DYKDDDDK tag antibody ECS epitope tag antibody ECS tag antibody Enterokinase Cleavage Site epitope tag antibody Enterokinase Cleavage Site tag antibody FLAG antibody FLAG tag antibody |
IHC
Immunohistochemical analysis of paraffin-embedded HeLa transfected with FLAG-tagged Claudin 18.2 (C-terminal) cells with Rabbit anti-DYKDDDDK Tag (FLAG) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling DYKDDDDK Tag (FLAG) with Rabbit anti-DYKDDDDK Tag (FLAG) antibody at 1/10,000 dilution. HeLa cells, transfected with FLAG-tagged empty control, Claudin 18.2 (C-terminal) or Histone H3.1 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-DYKDDDDK Tag (FLAG) antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells transfected with FLAG-tagged Otx1 (C-terminal) labeling DYKDDDDK Tag (FLAG). Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
DYKDDDDK Tag (FLAG) was immunoprecipitated in 2µg L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate with Rabbit anti-DYKDDDDK Tag (FLAG) antibody. Western blot was performed from the immunoprecipitate using DYKDDDDK Tag (FLAG) at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/100,000 dilution was used for 1 hour at room temperature. Lane 1: L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate (input). Lane 2: Rabbit IgG instead of Rabbit anti-DYKDDDDK Tag (FLAG) antibody IP in L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate. Lane 3: Rabbit anti-DYKDDDDK Tag (FLAG) antibody IP in L-929 transfected with FLAG-tagged CD5 (C-terminal) cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 17 seconds| Application Notes | IHC:1:1000 ICC:1:10000 FC:1:10000 IP:1-2ug/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
