DDDDK tag antibody (Equivalent To FLAG® Antibodies)
DDDDK tag Polyclonal Antibody for IF, WB, ELISA
Host / Isotype
Rabbit / IgG
Reactivity
recombinant protein and More (2)
Applications
IF, WB, ELISA
Conjugate
Unconjugated
142
Cat no : 20543-1-AP
Synonyms
DDDDK tag, DYKDDDDK
Validation Data Gallery
Tested Applications
| Positive WB detected in | recombinant protein protein |
| Positive IF detected in | Transfected HEK-293 cells |
Recommended dilution
| Application | Dilution |
|---|---|
| Western Blot (WB) | WB : 1:200-1:1000 |
| Immunofluorescence (IF) | IF : 1:10-1:100 |
| Sample-dependent, check data in validation data gallery | |
Published Applications
| WB | See 33 publications below |
| IHC | See 1 publications below |
| IF | See 7 publications below |
| IP | See 6 publications below |
| FC | See 1 publications below |
Product Information
The 20543-1-AP immunogen is Fusion Protein DDDDK tag protein expressed in E. coli.
| Reactivity | recombinant protein |
| Cited Species | human, mouse, recombinant protein |
| Host / Isotype | Rabbit / IgG |
| Class | Polyclonal |
| Type | Antibody |
| Immunogen | DDDDK tag fusion protein Ag2329 |
| Full Name | DDDDK tag |
| Calculated molecular weight | |
| Observed molecular weight | |
| GenBank accession number | |
| Gene symbol | |
| Gene ID (NCBI) | |
| Conjugate | Unconjugated |
| Form | Liquid |
| Purification Method | Antigen affinity purification |
| Storage Buffer | PBS with 0.1% sodium azide and 50% glycerol pH 7.3. |
| Storage Conditions | Store at -20oC. Aliquoting is unnecessary for -20oC storage. |
Background Information
Protein tags are protein or peptide sequences located either on the C- or N- terminal of the target protein, which facilitates one or several of the following characteristics: solubility, detection, purification, localization and expression. The DYKDDDDK(FLAG) peptide has been used extensively as a general tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion. N-terminal DDDDK vectors provide an Ek cleavage site for removal of the fusion tag. The DDDDK peptide is likely to be located on the surface of a fusion protein because of its hydrophilic nature. As a result, the DDDDK peptide is more likely to be accessible to antibodies. A DDDDK-tag can be used in many different assays that require recognition by an antibody, such as western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization and so on. This antibody is a rabbit polyclonal antibody raised against 3xFlag (3xDYKDDDDKT) sequence and recognizes the (1x) and (3x)DYKDDDDK peptide and detects DDDDK-tagged proteins. Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.
Protocols
| Product Specific Protocols | |
|---|---|
| WB protocol for DDDDK tag antibody 20543-1-AP | Download protocol |
| IF protocol for DDDDK tag antibody 20543-1-AP | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
| Species | Application | Title |
|---|---|---|
Front Immunol Long Interleukin-22 Binding Protein Isoform-1 Is an Intracellular Activator of the Unfolded Protein Response. | ||
Mol Immunol CpG oligodeoxynucleotide-specific duck TLR21 mediates activation of NF-κB signaling pathway and plays an important role in the host defence of DPV infection. | ||
Viruses PSMB1 Negatively Regulates the Innate Antiviral Immunity by Facilitating Degradation of IKK-ε. | ||
Cell Death Differ Glutathione S-transferases P1 protects breast cancer cell from adriamycin-induced cell death through promoting autophagy. | ||
Biochem Pharmacol The nuclear receptor Shp regulates morphine withdrawal syndrome via modulation of Ugt2b expression in mice. | ||
Free Radic Biol Med SIRT5-mediated SDHA desuccinylation promotes clear cell renal cell carcinoma tumorigenesis. |
Reviews
The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.
FH Dipen Rajgor (11-11-2019) | Great antibody for Western blotting. I used it at 1:1000 and got a super strong signal. Will use at a lower concentration next time.
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FH Sara Parker (10-24-2019) | Works really well for both IP of overexpressed FLAG-tagged proteins and WB!
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FH Nikhil Ghate (10-16-2019) | Flag tagged p53 expressed in H1299 cells.
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FH Shivani Gupta (09-26-2019) | Works better than another brand I had tested.
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FH Corey Goyeneche (08-23-2019) | This antibody has worked well for western blotting as well as immunoprecipitation. The lots have been consistent so far, we will definitely be ordering more.
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FH Kishor Pant (06-23-2019) | Works very good for WB at 1:1000 and IP at 1:200.
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FH David Timm (06-19-2019) | This is working very well for me. I ran out of another brand and got this one because I saw you had it and it was a better value. After testing it in the lab, it works just as well, if not better than the previous one. A very strong signal in my experiments.
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FH Lana Yablonska (04-20-2019) | SDS-PAGE: 10 ug/ul RIPA lysate of whole cells 4-12% Bis-tris gradient gel Transfer: Immobilon-FL transfer membranes (Millipore) O/N at 30V, 4C Blocking: SEA Block Blocking Buffer 1h Primary Ab: DDDDK pRb Cat# 20543-1-AP O/N incubation at 4C Secondary Ab: IRDye 800CW Goat anti-Rabbit 1 h incubation at room temperature Lines on WB: 1. BioRad Precision Plus Protein standards 2. Whole cell lysate of HEK293t cells (negative transfection) 3. Whole cell lysate of HEK293t cells transfected with FLAG-tag construct 27 kDa 3. Whole cell lysate of HEK293t cells transfected with FLAG-tag construct 40 kDa
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FH Luce Perie (11-12-2018) |
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