DYKDDDDK Epitope Tag Antibody (1042E) [Unconjugated]

Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with C-terminal DYKDDDDK-tagged Abro1, N-terminal DYKDDDDK-tagged UCHL5, and N-terminal DYKDDDDK-tagged RNF20. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-DYKDDDDK Epitope Tag Monoclonal Antibody (Catalog # MAB8529) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific bands were detected for DYKDDDDK-tagged proteins at approximately 55 kDa, 40 kDa, 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of HEK293 human embryonic kidney cell line transfected with C-terminal DYKDDDDK-tagged Abro1 and N-terminal DYKDDDDK-tagged RNF20, loaded at 0.1 mg/mL. Specific band were detected for DYKDDDDK-tagged proteins at approximately 64 and 136 kDa (as indicated) using 1 µg/mL of Rabbit Anti-DYKDDDDK Epitope Tag Monoclonal Antibody (Catalog # MAB8529). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

HEK293 cells transfected with DYKDDDDK-tagged proteins (filled histogram) vs Irrelevant HEK293 wild type cells (open histogram) were stained with Rabbit Anti-DYKDDDDK Epitope Tag Monoclonal Antibody (Catalog # MAB8529) at 0.001 ug/106 cells followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed and permeabilized with Flow Cytometry Fixation Buffer (Catalog # FC004). View our protocol for Staining Intracellular Molecules.

• Reactivity: All-Multi
• Applications: WB, Simple Western, IP, Flow
• Clone: 1042E
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

DYKDDDDK Epitope Tag Antibody (1042E) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: DYKDDDDK epitope tag
• Specificity: Detects N-terminal and C-terminal DYKDDDDK (FLAG^®) epitope tagged proteins in Western blot.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunoprecipitation 1-4 ug/100 ug cell lysate
  • Intracellular Staining by Flow Cytometry 0.001 ug/10^6 cells
  • Simple Western 1 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Preservative: Sodium Azide
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for DYKDDDDK Epitope Tag Antibody (1042E) [Unconjugated]

• DDDDK epitope tag
• DDK
• DYKDDDDK Epitope Tag
• DYKDDDDK tag
• ECS epitope tag
• ECS tag
• Enterokinase Cleavage Site epitope tag
• Enterokinase Cleavage Site tag
• epitope tag flag
• FLAG (FLAG is a trademark of Sigma Aldrich)
• FLAG Tag
• Ty 1 Tag
• xxxDDDDK epitope tag

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.

Publications for DYKDDDDK Epitope Tag Antibody (MAB8529)(5)

Publications using MAB8529

• Santorelli, M;Bhamidipati, PS;Courte, J;Swedlund, B;Jain, N;Poon, K;Schildknecht, D;Kavanagh, A;MacKrell, VA;Sondkar, T;Malaguti, M;Quadrato, G;Lowell, S;Thomson, M;Morsut, L; Control of spatio-temporal patterning via cell growth in a multicellular synthetic gene circuit Nature communications 2024-11-19 [PMID: 39562554] (Flow Cytometry, Mouse)
• PR Kamble, SR Patkar, AA Breed, BR Pathak N-glycosylation status of Trop2 impacts its surface density, interaction with claudin-7 and exosomal release Archives of biochemistry and biophysics, 2021-11-10;714(0):109084. 2021-11-10 [PMID: 34774484] (Western Blot, Human)
• WH Liu, K Völse, D Senft, I Jeremias A reporter system for enriching CRISPR/Cas9 knockout cells in technically challenging settings like patient models Scientific Reports, 2021-06-16;11(1):12649. 2021-06-16 [PMID: 34135367] (Western Blot, Human)
• W Zhang, G Yin, H Zhao, H Ling, Z Xie, C Xiao, Y Chen, Y Lin, T Jiang, S Jin, J Wang, X Yang Secreted KIAA1199 promotes the progression of rheumatoid arthritis by mediating hyaluronic acid degradation in an ANXA1-dependent manner Cell Death & Disease, 2021-01-20;12(1):102. 2021-01-20 [PMID: 33473125] (Immunoprecipitation, Human)
• Li YL, Cai WF, Wang L et al. Identification of the Functional Autophagy-Regulatory Domain in HCLS1-Associated Protein X-1 That Resists Against Oxidative Stress. DNA Cell Biol 5/1/2018 [PMID: 29461873]

Reviews for DYKDDDDK Epitope Tag Antibody (MAB8529) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Verified Customer ICC Human 09/29/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: HEK293 cell line
• Species: Human
• Lot: CIZU0115011

Comments

• Comments: NOVUSLOOKSLIKE

reviewed by: Verified Customer ICC Human 05/22/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: HEK293
• Species: Human
• Lot: CIZU0115011

FAQs for DYKDDDDK Epitope Tag Antibody (MAB8529). (Showing 1 - 5 of 5 FAQs).

1. 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.
2. What the theoretical molecular weight for FLAG Tag antibodies?
   • The TMW for DYKDDDDK epitope FLAG Tag is approximately 1.01 kDa.
3. 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.
4. 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.
5. 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.