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DYKDDDDK Tag (Anti-FLAG) Antibody Agarose Conjugated
Catalog Number: orb344560
| Catalog Number | orb344560 |
|---|---|
| Category | Antibodies |
| Description | Mouse monoclonal antibody against DYKDDDDK (FLAG tag) conjugated to Agarose. |
| Clonality | Monoclonal |
| Species/Host | Mouse |
| Isotype | IgG2a |
| Conjugation | Agarose |
| Form/Appearance | Suspension of agarose beads |
| Concentration | 0.99 mg |
| Buffer/Preservatives | 0.01% (w/v) Sodium Azide |
| Purity | Anti-DYKDDDDK Affinity Gel is a purified mouse IgG2a monoclonal antibody coupled to activated agarose. This product is intended for purification of proteins containing the FLAG epitope tag sequence. Binding Specificity: Anti-DYKDDDDK Affinity Gel binds the FLAG epitope tag sequence (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) fused to the amino terminal, carboxy terminal or internal locations of targeted recombinant proteins expressed in transfected or transformed cells. D-Y-K-D-D-D-D-K peptide (p/n 000-000-383) is recommended for competitive elution to recover fusion protein (see protocol). |
| Immunogen | Anti-DYKDDDDK Affinity Gel antibody was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH. |
| Tested applications | IP, WB |
| Dilution range | IP: 10 µL resin binds >1 µg FLAG fusion protein |
| Application notes | Anti-DYKDDDDK Affinity Gel has been tested by SDS-Page, IP, and western blot and is optimally suited for immunoprecipitation and purification of FLAG tagged fusion proteins. Anti-DYKDDDDK Affinity Gel antibody recognizes the FLAG epitope tag fused to either the amino- or carboxy- terminal ends or an internal location of targeted fusion proteins. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. DYKDDDDK is the most commonly used hydrophilic octapeptide tag. Use D-Y-K-D-D-D-D-K peptide (p/n 000-000-383) for competitive elution to recover fusion protein (see protocol). Anti-FLAG is a registered trademark of Sigma-Aldrich. Refer to the Protocol for complete instructions for use including preferred buffers for elution. Do not use buffers that may denature the anti-DYKDDDDK antibody. |
| Antibody Type | Primary Antibody |
| Clone Number | 29E4.G7 |
| Storage | Store vial at 4°C prior to opening. |
| Alternative names | mouse anti-FLAG tag agarose gel, mouse anti-DYKDDD Read more... |
| Note | For research use only |
SDS-PAGE of Anti-DYKDDDDK (FLAG® tag) Affinity Gel. Lane 1: Cell lysate before purification. Lane 2: Flow through (used cell lysate). Lane 3: Purified DYKDDDDK (FLAG® tag) recombinant protein (arrowhead). Load: (6 µl per lane). Predicted/Observed size: 70kDa for DYKDDDDK tagged recombinant protein.
SDS-PAGE of Anti-DYKDDDDK (FLAG® tag) Affinity Gel. Lane 1: crude lysate containing over-expressed DYKDDDK-tagged recombinant protein. Lane 2: unbound flow-through of consisting of endogenous E.coli proteins. Lane 3: enriched recombinant protein. Load: 5 µg protein, 15 µl flow through. Predicted/Observed size: 70kDa for DYKDDDDK tagged recombinant protein.
SDS-PAGE of Anti-DYKDDDDK (FLAG® tag) Affinity Gel. Lane 1: DYKDDDDK tagged recombinant protein. Lane 2: flow through. Lane 3: wash. Lane 4: eluted fraction one. Lane 5: eluted fraction two. Lane 6: eluted fraction three. Lane 7: blank. Lane 8: molecular weight markers. Load: 5 µg protein lane 1, 15 µl per each other lane. Predicted/Observed size: 135kDa for DYKDDDDK tagged recombinant protein.
