DYKDDDDK Epitope Tag Antibody (OTI4C5)
Novus Biologicals, part of Bio-Techne | Catalog # NBP1-71705
Key Product Details
Species Reactivity
Non-species specific
Applications
Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Simple Western, Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # OTI4C5
Concentration
Please see the vial label for concentration. If unlisted please contact technical services.
Product Specifications
Immunogen
Anti-DDK monoclonal antibody is produced by immunizing mice with a synthetic peptide (DYKDDDDK) coupled to KLH.
Reactivity Notes
DDK epitope tag (DYKDDDDK)
Specificity
The anti-DDK (also known as Flag (R) ) antibody recognizes over expressed recombinant proteins containing the DDK epitope tag (DYKDDDDK) fused to either the amino- or carboxy-termini of targeted proteins in transfected mammalian cells or other expression systems.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Applications for DYKDDDDK Epitope Tag Antibody (OTI4C5)
Application
Recommended Usage
Flow Cytometry
1:1000
Immunocytochemistry/ Immunofluorescence
1:1000
Immunoprecipitation
1:200
Simple Western
1:250
Western Blot
1:2000
Application Notes
Only recognizes n-terminal form in Simple Western. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Formulation, Preparation, and Storage
Purification
Protein A or G purified
Formulation
PBS (pH 7.3), 1.0% BSA and 50% Glycerol
Preservative
0.02% Sodium Azide
Concentration
Please see the vial label for concentration. If unlisted please contact technical services.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: DYKDDDDK Epitope Tag
The epitope tag, DYKDDDDK, is comprised of an eight amino acid sequence derived from the gene-10 product from bacteriophage T7 and contains an enterokinase-cleavage site (1). Its hydrophilic nature and short peptide sequence reduce possible interference with protein expression, post-translational modifications, protein interactions, and protein function (2). The DYKDDDDK (FLAG(R)) epitope tag can either be fused to the N-terminus, C-terminus, or placed at an internal position of the target protein, facilitating characterization of the protein of interest using immunodetection methods. Compared to the standard DYKDDDDK epitope tag antibody, the rat monoclonal DYKDDDDK Epitope Tag antibody (clone L5) is 2-8 times more sensitive (3).
/>References
/>1. Brizzard BL, Chubet RG, Vizard DL. (1994) Immunoaffinity purification of (FLAG(R)) epitope-tagged bacterial alkaline phosphatase using a novel monoclonal antibody and peptide elution. Biotechniques. 16(4):730-5. PMID: 8024796
2. Einhauer A, Jungbauer A. (2001) The (FLAG(R)) peptide, a versatile fusion tag for the purification of recombinant proteins. J Biochem Biophys Methods. 49(1-3):455-65. PMID: 11694294
3. Park SH, Cheong C, Idoyaga J, Kim JY, Choi JH, Do Y, Lee H, Jo JH, Oh YS, Im W, Steinman RM, Park CG. (2008) Generation and application of new rat monoclonal antibodies against synthetic DYKDDDDK Epitope Tag (FLAG(R)) and OLLAS tags for improved immunodetection. J Immunol Methods. 331(1-2):27-38. PMID: 18054954
FLAG(R) is a trademark of Sigma-Aldrich.
Alternate Names
FLAG Tag
Additional DYKDDDDK Epitope Tag Products
Product Documents for DYKDDDDK Epitope Tag Antibody (OTI4C5)
Product Specific Notices for DYKDDDDK Epitope Tag Antibody (OTI4C5)
FLAG(R) and ANTI-FLAG(R) are registered trademarks of Sigma-Aldrich Co. LLC.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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