|Positive WB detected in||recombinant protein protein|
|Positive IF detected in||Transfected HEK-293 cells|
|Western Blot (WB)||WB : 1:200-1:1000|
|Immunofluorescence (IF)||IF : 1:10-1:100|
|Sample-dependent, check data in validation data gallery|
The 20543-1-AP immunogen is Fusion Protein DDDDK tag protein expressed in E. coli.
|Cited Species||human, mouse, recombinant protein|
|Host / Isotype||Rabbit / IgG|
|Immunogen||DDDDK tag fusion protein Ag2329|
|Full Name||DDDDK tag|
|Calculated molecular weight|
|Observed molecular weight|
|GenBank accession number|
|Gene ID (NCBI)|
|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.|
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.
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The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.
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.
Sara Parker (10-24-2019)
Works really well for both IP of overexpressed FLAG-tagged proteins and WB!
Nikhil Ghate (10-16-2019)
Flag tagged p53 expressed in H1299 cells.
Shivani Gupta (09-26-2019)
Works better than another brand I had tested.
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.
Kishor Pant (06-23-2019)
Works very good for WB at 1:1000 and IP at 1:200.
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.
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
Luce Perie (11-12-2018)