DYKDDDDK Epitope Tag Antibody [Alexa Fluor® 532] Summary
| Description |
This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet. |
| Immunogen |
To produce DYKDDDDK Epitope Tag Antibody, Rabbits were immunized with ECS enterokinase cleavage site (xxxDDDDK) conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support. |
| Specificity |
DYKDDDDK Epitope Tag Antibody reacts with FLAG. |
| Isotype |
IgG |
| Clonality |
Polyclonal |
| Host |
Rabbit |
| Purity |
Immunogen affinity purified |
| Innovator's Reward |
Test in a species/application not listed above to receive a full credit towards a future purchase. |
Applications/Dilutions
| Dilutions |
- ELISA
- Immunohistochemistry
- Western Blot
|
| Application Notes |
Optimal dilution of this antibody should be experimentally determined. |
| Theoretical MW |
1.01 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
Packaging, Storage & Formulations
| Storage |
Store at 4C in the dark. |
| Buffer |
50mM Sodium Borate |
| Preservative |
0.05% Sodium Azide |
| Purity |
Immunogen affinity purified |
Notes
FLAG(R) and ANTI-FLAG(R) are registered trademarks of Sigma-Aldrich Co. LLC.
Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.
Alternate Names for DYKDDDDK Epitope Tag Antibody [Alexa Fluor® 532]
Background
Fusion proteins, in which a known peptide such as a
His-Tag,
GST-Tag, or
DYKDDDDK (FLAG(R)) is cloned with the target of interest, were developed to simplify protein isolation. Since specific antibodies may not be available against the protein of interest,
epitope tag antibodies are commonly used to detect protein expression.
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.
Limitations
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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Product General Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
Video Protocols
FAQs for DYKDDDDK Epitope Tag Antibody (NB600-345AF532). (Showing 1 - 5 of 5 FAQ).
-
Are the FLAG Tag antibodies validated in Simple Western?
- Yes, we offer 3 FLAG Tag antibodies that have been tested in Simple Western: NBP1-71705, NBP1-97410, and MAB8529.
-
What the theoretical molecular weight for FLAG Tag antibodies?
- The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.
-
Will FLAG Tag antibodies detect internal FLAG tags? My FLAG sequence is sandwiched between two other protein sequences.
- If you are applying your sample(s) to SDS-PAGE, under reducing condition and follow by Western Blot, DYKDDDDK epitope tag antibodies should be able to detect the tag region located internally. However, it depends on the property of your chimeric protein and if your tag is in the hydrophobic or hydrophilic region. Although DYKDDDDK is very hydrophilic, it may not be exposed to the surface if embedded deeply inside the protein with a high content of hydrophobic amino acids with or without covalently-linked modifications such as lipid moieties.
-
Will FLAG Tag antibodies recognize FLAG at the C-terminal end?
- Yes, FLAG Tag antibodies can recognize FLAG-tag at both the N- or C-terminal.
-
Do FLAG Tag antibodies comes in lyophilized form?
- The following FLAG Tag antibodies come in lyophilized form: NBP1-97393, NBP1-97401, NBP1-97399, NBP1-97400, and NBP1-97402.
![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
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