A6001-25mg | 3X (DYKDDDDK) Peptide [402750-12-3] Clinisciences

Background

The DYKDDDDK-tag system utilizes a short, hydrophilic 8- amino-acid peptide that is fused to the protein of interest1. The DYKDDDDK peptide binds to the antibody M1. Whether binding is calcium-dependent manner2 or –independent3 remains controversial. A disadvantage of the system is that the monoclonalantibody purification matrix is not as stable as others. In general, small tags can be detected with specific monoclonal antibodies.

To improve the detection of the DYKDDDDK tag the 3x (DYKDDDDK) system has been developed. This threetandem DYKDDDDK epitope is hydrophilic, 22-amino-acids long and can detect up to 10 fmol of expressed fusion protein. The DYKDDDDK-tagged maltodextrin-binding protein of Pyrococcus furiosus has been crystallized4 and the quality of the crystals was very similar to that of crystals of untagged protein.

Finally, the DYKDDDDK-tag can be removed by treatment with enterokinase, which is specific for the five C-terminal amino acids of the peptide sequence5.

References:
1. Hopp TP, Prickett KS, VL, Libby RT, March CJ, Ceretti DP, Urdal DL, Conlon PJ (1988) A short polypeptide marker sequence useful for recombinant protein identification and purification. Bio/Technology 6:1204–1210.
2. Hopp TP, Gallis B, Prikett KS (1996) Metal-binding properties of a calcium dependent monoclonal antibody. Mol Immunol 33:601–608.
3. Einhauer A, Jungbauer A (2000) Kinetics and thermodynamical properties of the monoclonal antibody M1 directed against the FLAG peptide. 20th International symposium on the separation of proteins, peptides, and polynucleotides (ISPPP). Lublijana, Slovenia, November 5–8, 2000.
4. Bucher MH, Evdokimov AG, Waugh DS (2002) Differential effects of short affinity tags on the crystallization of Pyrococcus furiosus maltodextrin-binding protein. Biol Cryst 58:392–397.
5. Maroux S, Baratti J, Desnuelle P (1971) Purification and specificity of procine enterokinase. J Biol Chem 246:5031–5039.

Product Citation

1. Yao Gong, Gaurav Behera, et al. "HypDB: A functionally annotated web-based database of the proline hydroxylation proteome." PLoS Biol. 2022 Aug 26;20(8):e3001757. PMID: 36026437
2. Daniel Frank, Sarah E Garnish, et al. "Ubiquitylation of RIPK3 beyond-the-RHIM can limit RIPK3 activity and cell death." iScience. 2022 Jun 17;25(7):104632. PMID: 35800780
3. Md Mahfuz Al Mamun, Muhammad Riaz Khan, et al. "Stub1 maintains proteostasis of master transcription factors in embryonic stem cells." Cell Rep. 2022 Jun 7;39(10):110919. PMID: 35675767
4. Yang Xu, Han Han, et al. "Active conformation of the p97-p47 unfoldase complex." Nat Commun. 2022 May 12;13(1):2640. PMID: 35552390
5. Lin Liu, Jarrod J. Sandow, et al. "Tankyrase-mediated ADP-ribosylation is a regulator of TNF-induced death." Sci Adv. 2022 May 13;8(19):eabh2332. PMID: 35544574
6. Michael G Hanna, Patreece H Suen, et al. "SHIP164 is a chorein motif lipid transfer protein that controls endosome–Golgi membrane traffic." J Cell Biol. 2022 Jun 6;221(6):e202111018. PMID: 35499567
7. Zhongxia Yi, René M Arvola, et al. "Mammalian UPF3A and UPF3B can activate nonsense‐mediated mRNA decay independently of their exon junction complex binding." EMBO J. 2022 May 16;41(10):e109202. PMID: 35451102
8. Haining Zhou, Chad B Stein, et al. "Rixosomal RNA degradation contributes to silencing of Polycomb target genes." Nature. 2022 Apr;604(7904):167-174. PMID: 35355014
9. Akhil Gargey, Yuri E Nesmelov. "The Local Environment of Loop Switch 1 Modulates the Rate of ATP-Induced Dissociation of Human Cardiac Actomyosin." Int J Mol Sci. 2022 Jan 22;23(3):1220. PMID: 35163146
10. Yi Zhao, Pierre A. Miranda Herrera, et al. "Function-guided proximity mapping unveils electrophilic-metabolite sensing by proteins not present in their canonical locales." Proc Natl Acad Sci U S A. 2022 Feb 1;119(5):e2120687119. PMID: 35082156
11. Mutchler, Ashley S. "The role of Esc2 and DNA in Smc5/6-mediated Sgs1-Top3-Rmi1 sumoylation." Texas State University. 2022.
12. Connor Quinn. "Molecular Drivers of Disease Pathogenesis in Nonalcoholic Steatohepatitis." Temple University. August 2022.
13. Frank, Daniel, et al. "The role of RIPK3 ubiquitylation and MLKL signalling during cell death and autophagy." University of Melbourne. 2021.
14. Jessica J Lenoir, Jean-Patrick Parisien, et al. "Immune regulator LGP2 targets Ubc13/UBE2N to mediate widespread interference with K63 polyubiquitination and NF-κB activation." Cell Rep. 2021 Dec 28;37(13):110175. PMID: 34965427
15. Wei Yuan, Qais Al-Hadid, et al. "TDRD3 promotes DHX9 chromatin recruitment and R-loop resolution." Nucleic Acids Res. 2021 Sep 7;49(15):8573-8591. PMID: 34329467
16. Xingyu Pan, Jingrong Zhao, et al. "5'-UTR SNP of FGF13 causes translational defect and intellectual disability." Elife. 2021 Jun 29;10:e63021. PMID: 34184986
17. Daniel L. Kober, Arun Radhakrishnan, et al. "Scap structures highlight key role for rotation of intertwined luminal loops in cholesterol sensing." Cell. 2021 Jul 8;184(14):3689-3701.e22. PMID: 34139175
18. Tingjuan Deng, Boli Hu, et al. "Inhibition of Antiviral Innate Immunity by Avibirnavirus VP3 via Blocking TBK1-TRAF3 Complex Formation and IRF3 Activation." mSystems. 2021 May 11;6(3):e00016-21. PMID: 34055057
19. Patricia R Nano, Taylor K Johnson, et al. "Structure-activity mapping of ARHGAP36 reveals regulatory roles for its GAP homology and C-terminal domains." PLoS One. 2021 May 17;16(5):e0251684. PMID: 33999959
20. Lingli He, Wentao Yu, et al. "An optimized two-step chromatin immunoprecipitation protocol to quantify the associations of two separate proteins and their common target DNA." STAR Protoc. 2021 Apr 27;2(2):100504. PMID: 33997818
21. Yevgeni Yegorov, Eleonora Sendersky, et al. "A Cyanobacterial Component Required for Pilus Biogenesis Affects the Exoproteome." mBio. 2021 Mar 16;12(2):e03674-20. PMID: 33727363
22. Ke Zhang, Lisa Miorin, et al. "Nsp1 protein of SARS-CoV-2 disrupts the mRNA export machinery to inhibit host gene expression." Sci Adv. 2021 Feb 5;7(6):eabe7386. PMID: 33547084
23. Haijing Guo, Jen-Hsuan Wei, et al. "Importin α phosphorylation promotes TPX2 activation by GM130 to control astral microtubules and spindle orientation." J Cell Sci. 2021 Feb 19;134(4):jcs258356. PMID: 33526712
24. Joshua A Lees, PeiQi Li, et al. "Insights into Lysosomal PI (3, 5) P2 Homeostasis from a Structural-Biochemical Analysis of the PIKfyve Lipid Kinase Complex." Mol Cell. 2020 Nov 19;80(4):736-743.e4. PMID: 33098764
25. Akhil Gargey, Shiril Bhardwaj Iragavarapu, et al. "Electrostatic interactions in the SH1-SH2 helix of human cardiac myosin modulate the time of strong actomyosin binding." J Muscle Res Cell Motil. 2021 Jun;42(2):137-147. PMID: 32929610
26. Corinna Kosnopfel, Tobias Sinnberg, et al. "Tumour Progression Stage-Dependent Secretion of YB-1 Stimulates Melanoma Cell Migration and Invasion." Cancers (Basel) 2020 Aug 18;12(8):2328. PMID: 32824741
27. Martin Mehnert, Rodolfo Ciuffa, et al. "Multi-layered proteomic analyses decode compositional and functional effects of cancer mutations on kinase complexes." Nat Commun. 2020 Jul 16;11(1):3563. PMID: 32678104
28. Wu Y, Jin S, et al. "Selective autophagy controls the stability of transcription factor IRF3 to balance type I interferon production and immune suppression." Autophagy. 2020;1-14. PMID: 32476569
29. Howard SM, Ceppi I, et al. "The internal region of CtIP negatively regulates DNA end resection." Nucleic Acids Res. 2020;48(10):5485-5498. PMID: 32347940
30. Jesse R. Poganik, Alexandra K. Van Hall-Beauvais, et al. "The mRNA-binding protein HuR is a kinetically-privileged electrophile sensor." Helvetica Chimica Acta.10.1002.
31. Haijing Guo, Jen-Hsuan Wei, et al. "TPX2 activation by GM130 controls astral microtubule formation and spindle orientation." bioRxiv. March 17, 2020.
32. Xiao J, Li W, et al. "Targeting 7-Dehydrocholesterol Reductase Integrates Cholesterol Metabolism and IRF3 Activation to Eliminate Infection." Immunity. 2020;52(1):109–122.e6. PMID: 31882361
33. Andrii Bugai. "REGULATION OF POL II TRANSCRIPTIONAL RESPONSE TO DNA DAMAGE BY RBM7 AND P-TEFb." University of Helsinki.2020.
34. Benjamin Micah Milrod Grant. "Calmodulin Disrupts Plasma Membrane Localization of Farnesylated KRAS4b by Sequestering its Lipid Moiety." University of Toronto. 2019.
35. Liu J, Feng J, et al. "Arginine methylation-dependent LSD1 stability promotes invasion and metastasis of breast cancer." EMBO Rep. 2019 Dec 12:e48597. PMID: 31833203
36. Luo A, Chen Y. "Label-free Interactome Analysis Revealed an Essential Role of CUL3-KEAP1 Complex in Mediating the Ubiquitination and Degradation of PHD2." J Proteome Res. 2019 Nov 25. PMID: 31763849
37. Peter Kindgren, Maxim Ivanov, et al. "Native Elongation Transcript sequencing reveals temperature dependent dynamics of nascent RNAPII transcription in Arabidopsis." bioRxiv. 2019 November 08.
38. Qin X, Tang HT, et al. "An Orthogonal Tyrosyl-tRNA Synthetase/tRNA Pair from a Thermophilic Bacterium for an Expanded Eukaryotic Genetic Code." Biochemistry. 2019 Nov 8. PMID: 31703481
39. Wang X, Xu P, et al. "Cellular Cleavage and Polyadenylation Specificity Factor 6 (CPSF6) Mediates Nuclear Import of Human Bocavirus 1 NP1 Protein and Modulates Viral Capsid Protein Expression." J Virol. 2019 Oct 30. pii: JVI.01444-19. PMID: 31666379
40. Kumar Singh A, Tamrakar A, et al. "Splicing regulator SRSF1-3 that controls somatic hypermutation of IgV genes interacts with topoisomerase 1 and AID." Mol Immunol. 2019 Oct 14;116:63-72. PMID: 31622795
41. Chen RX, Chen X, et al. "N(6)-methyladenosine modification of circNSUN2 facilitates cytoplasmic export and stabilizes HMGA2 to promote colorectal liver metastasis." Nat Commun. 2019 Oct 16;10(1):4695. PMID: 31619685
42. Chavez C, Cruz-Becerra G, et al. "The tardigrade damage suppressor protein binds to nucleosomes and protects DNA from hydroxyl radicals." Elife. 2019 Oct 1;8. pii: e47682. PMID: 31571581
43. Zhang J, Ali AM, et al. "Disease-Causing Mutations in SF3B1 Alter Splicing by Disrupting Interaction with SUGP1." Mol Cell. 2019 Aug 9. pii: S1097-2765(19)30547-7. PMID: 31474574
44. Elizabeth Anne Moore. "Discovering and Drugging Anti-Cancer Targets of Natural Compounds Using Chemoproteomics." University of California. 2019.
45. Charles A. Berdan. "Utilizing Chemoproteomics to Discover Novel Druggable Hotspots Targeted by Natural Products." University of California. 2019.
46. Yingqi Liu, Xiaoqing Liu, et al. "The PRMT7-dependent methylation of shank2 modulates invasion-proliferation switching during breast cancer metastasis." bioRxiv. 2019 August 07.
47. Zhao Z, Wang L, et al. "β-Catenin/Tcf7l2-dependent transcriptional regulation of GLUT1 gene expression by Zic family proteins in colon cancer." Sci Adv. 2019 Jul 31;5 (7):eaax0698. PMID: 31392276
48. Valerie T. Ressler. "Endogenous and Chemical Modifications of Model Proteins." Massachusetts Institute of Technology. 2019.
49. Njomen E, Tepe JJ. "Regulation of Autophagic Flux by the 20S Proteasome." Cell Chem Biol. 2019 Jul 16. pii: S2451-9456(19)30210-7. PMID: 31327703
50. Ngaio Charlotte Smith. "Investigating the role of protein-protein and protein-DNA interactions in the function of Isl1." University of Sydney. 2019.
51. Piunti A, Smith ER, et al. "CATACOMB: An endogenous inducible gene that antagonizes H3K27 methylation activity of Polycomb repressive complex 2 via an H3K27M-like mechanism." Sci Adv. 2019 Jul 3;5(7):eaax2887. PMID: 31281901
52. Jasleen Singh, Vibhor Mishra, et al. "Reaction mechanisms of Pol IV, RDR2 and DCL3 drive RNA channeling in the siRNA-directed DNA methylation pathway." bioRxiv. 2019 June 22.
53. Dylan Charles Mitchell. "Development of a Chemoproteomic Approach to Study Kinase-Substrate Interactions and the Discovery of CDK4 as a 4E-BP1 Kinase." University of Michigan. 2019.
54. Miller CJ, Lou HJ, et al. "Comprehensive profiling of the STE20 kinase family defines features essential for selective substrate targeting and signaling output." PLoS Biol. 2019 Mar 21;17(3):e2006540. PMID: 30897078
55. Zhang M, Ergin V, et al. "Axonogenesis Is Coordinated by Neuron-Specific Alternative Splicing Programming and Splicing Regulator PTBP2." Neuron. 2019 Feb 20;101(4):690-706.e10. PMID: 30733148
56. Charles A Berdan, Raymond Ho, et al. "Parthenolide Covalently Targets and Inhibits Focal Adhesion Kinase in Breast Cancer Cells." bioRxiv. 2019 February 14.
57. Gomes ID, Pflum MKH. "Optimal substrate trapping mutants to discover substrates of HDAC1." Chembiochem. 2019 Jan 30. PMID: 30701667
58. Liang Z, Liang F, et al. "Binding of FANCI-FANCD2 Complex to RNA and R-Loops Stimulates Robust FANCD2 Monoubiquitination." Cell Rep. 2019 Jan 15;26(3):564-572.e5. PMID: 30650351
59. Bert van de Kooij, Pau Creixell, et al. "Comprehensive substrate specificity profiling of the human nek kinome reveals unexpected signaling outputs." bioRxiv. 2019 January 08
60. Roth L, Wakim J, et al. "Phosphorylation of the phosphatase PTPROt at Tyr(399) is a molecular switch that controls osteoclast activity and bone mass in vivo." Sci Signal. 2019 Jan 8;12(563). pii: eaau0240. PMID: 30622194
61. Sluder IT, Nitika, et al. "The Hsp70 co-chaperone Ydj1/HDJ2 regulates ribonucleotide reductase activity." PLoS Genet. 2018 Nov 19;14(11):e1007462. PMID: 30452489
62. Samant RS, Livingston CM, et al. "Distinct proteostasis circuits cooperate in nuclear and cytoplasmic protein quality control." Nature. 2018 Nov;563(7731):407-411. PMID: 30429547
63. Jessica N. Spradlin, Xirui Hu, et al. "Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation." bioRxiv. 2018 October 15.
64. JianWu, ShuangshuangNiu, et al. "Cryo-EM Structure of the Human Ribonuclease P Holoenzyme." Cell.Available online 25 October 2018.
65. Fu Y, Long MJC, et al. "Nuclear RNR-α antagonizes cell proliferation by directly inhibiting ZRANB3." Nat Chem Biol. 2018 Oct;14(10):943-954. PMID: 30150681
66. Ito T, Kasai Y, et al. "Quantitative Analysis of Interaction Between CADM1 and Its Binding Cell-Surface Proteins Using Surface Plasmon Resonance Imaging." Front Cell Dev Biol. 2018 Aug 7;6:86. PMID: 30131958
67. Andrii Bugai, Alexandre JC Quaresma, et al. "P-TEFb activation by RBM7 shapes a pro-survival transcriptional response to genotoxic stress." bioRxiv. 2018 August 17.
68. Lampert F, Stafa D, et al. "The multi-subunit GID/CTLH E3 ubiquitin ligase promotes cell proliferation and targets the transcription factor Hbp1 for degradation." Elife. 2018 Jun 18;7. pii: e35528. PMID: 29911972
69. Zi-Wei Chen, John R Bracamontes, et al. "Multiple Functional Neurosteroid Binding Sites on GABAA Receptors." bioRxiv. 2018 June 29.
70. Wu Z, Wang Y, et al."Ubiquitination of ABCE1 by NOT4 in Response to Mitochondrial Damage Links Co-translational Quality Control to PINK1-Directed Mitophagy." Cell Metab. 2018 Jul 3;28(1):130-144.e7. PMID: 29861391
71. Peter Kindgren, Ryan Ard, et al."Transcriptional read-through of the long non-coding RNA SVALKA governs plant cold acclimation." bioRxiv.2018 March 23
72. Caldas, et al. "Investigation of a transcription factor complex and intrinsically disordered proteins." The University of Sydney. 2018 February.
73. Hoshii T, Cifani P, et al. "A Non-catalytic Function of SETD1A Regulates Cyclin K and the DNA Damage Response." Cell. 2018 Feb 22;172(5):1007-1021.e17. PMID: 29474905
74. Anghel SA, McGilvray PT, et al. "Identification of Oxa1 Homologs Operating in the Eukaryotic Endoplasmic Reticulum." Cell Rep. 2017 Dec 26;21(13):3708-3716. PMID: 29281821
75. Wang X, Paucek RD, et al. "Molecular analysis of PRC2 recruitment to DNA in chromatin and its inhibition by RNA." Nat Struct Mol Biol. 2017 Dec;24(12):1028-1038. PMID: 29058709
76. Hsu M, Lue NF. "The mechanisms of K. lactis Cdc13 in telomere DNA-binding and telomerase regulation." DNA Repair (Amst). 2018 Jan;61:37-45. PMID: 29197718
77. Grossman EA, Ward CC, et al. "Covalent Ligand Discovery against Druggable Hotspots Targeted by Anti-cancer Natural Products." Cell Chem Biol. 2017 Sep 11. pii:S2451-9456(17)30311-2. PMID: 28919038
78. Li Y, Kim J. "Distinct roles of neuronal and microglial CB2 cannabinoid receptors in the mouse hippocampus." Neuroscience. 2017 Sep 6;363:11-25. PMID: 28888955
79. Wang WF, Yan L, et al. "HSP70-Hrd1 axis precludes the oncorepressor potential of N-terminal misfolded Blimp-1s in lymphoma cells." Nat Commun. 2017 Aug 25;8(1):363. PMID: 28842558
80. Ogami K, Richard P, et al. "An Mtr4/ZFC3H1 complex facilitates turnover of unstable nuclear RNAs to prevent their cytoplasmic transport and global translational repression." Genes Dev. 2017 Jun 15;31(12):1257-1271. PMID: 28733371
81. Ying Y, Wang XJ, et al. "Splicing Activation by Rbfox Requires Self-Aggregation through Its Tyrosine-Rich Domain." Cell. 2017 Jul 13;170(2):312-323.e10. PMID: 28708999
82. Wang Erickson AF, Deighan P, et al. "A novel RNA polymerase-binding protein that interacts with a sigma-factor docking site." Mol Microbiol. 2017 Jun 8. PMID: 28598017
83. Xiao X, Yang H, et al. "Structural determinants of APOBEC3B non-catalytic domain for molecular assembly and catalytic regulation." Nucleic Acids Res. 2017 May 30. PMID: 28575276
84.Tanu Talwar. "Biochemical Characterization of DDX43 (HAGE) Helicase." University of Saskatchewan. 2017 Mar.
85. Liu M, Feng Z, et al. "Tango1 spatially organizes ER exit sites to control ER export." J Cell Biol. 2017 Apr 3;216(4):1035-1049. PMID: 28280122
86. Zilio N, Boddy MN. "Improved Tandem Affinity Purification Tag and Methods for Isolation of Proteins and Protein Complexes from Schizosaccharomyces pombe. Cold Spring Harb Protoc." 2017 Mar 1;2017(3):pdb.prot091611. PMID: 28250214
87. Belyaeva OV, Adams MK, et al. "The antagonistically bifunctional retinoid oxidoreductase complex is required for maintenance of all-trans-retinoic acid homeostasis." J Biol Chem. 2017 Apr 7;292(14):5884-5897. PMID: 28232491
88. Zhang X, Smits AH, et al. "An Interaction Landscape of Ubiquitin Signaling. Mol Cell." 2017 Mar 2;65(5):941-955.e8. PMID: 28190767
89. Geng P, Zhang Y, et al. "Automethylation of protein arginine methyltransferase 7 and its impact on breast cancer progression." FASEB J. 2017 Feb 10. pii: fj.201601196R. PMID: 28188177
90. Davis AM, Ramirez J, et al. "Identification of influenza A nucleoprotein body domain residues essential for viral RNA expression expose antiviral target. " Virol J. 2017 Feb 7;14(1):22. PMID: 28173821
91. Nalawansha DA, Pflum MK. "LSD1 Substrate Binding and Gene Expression Are Affected by HDAC1-Mediated Deacetylation." ACS Chem Biol. 2017 Jan 20;12(1):254-264. PMID: 27977115
92. Manthey GM, Clear AD, et al. "Homologous recombination in budding yeast expressing the human RAD52 gene reveals a Rad51-independent mechanism of conservative double-strand break repair." Nucleic Acids Res. 2017 Feb 28;45(4):1879-1888. PMID: 27923995
93. Hoang TT, Raines RT. "Molecular basis for the autonomous promotion of cell proliferation by angiogenin." Nucleic Acids Res. 2017 Jan 25;45(2):818-831. PMID: 27915233
94. Kazazian K, Go C, et al. "Plk4 Promotes Cancer Invasion and Metastasis through Arp2/3 Complex Regulation of the Actin Cytoskeleton." Cancer Res. 2017 Jan 15;77(2):434-447. PMID: 27872092
95. Shen W, Deng X, et al. "Analysis of the Cis and Trans Requirements for DNA Replication at the Right End Hairpin of the Human Bocavirus 1 Genome." J Virol. 2016 Jun 22. PMID: 27334591
96. Hresko RC, Kraft TE, et al. "Mammalian Glucose Transporter Activity Is Dependent upon Anionic and Conical Phospholipids." J Biol Chem. 2016 Aug 12;291(33):17271-82. PMID: 27302065
97. Rambout X, Detiffe C, et al. "The transcription factor ERG recruits CCR4-NOT to control mRNA decay and mitotic progression." Nat Struct Mol Biol. 2016 Jul;23(7):663-72. PMID: 27273514
98. Low JK, Webb SR, et al. "CHD4 is a Peripheral Component of the Nucleosome Remodeling and Deacetylase Complex." J Biol Chem. 2016 May 27. PMID: 27235397
99. Alicia Morgan Davis. "Characterization of Influenza Nucleoprotein body domain as antiviral target." California State University.June 2016.
100. Paul W. Hruz, Chair,et al. "Modulation of Human and Malarial Glucose Transporter Activity by Lipids and Small Molecules." Washington University in St. Louis.May 2016.
101. Silva AP, Ryan DP, et al. "The N-terminal Region of Chromodomain Helicase DNA-binding Protein 4 (CHD4) Is Essential for Activity and Contains a High Mobility Group (HMG) Box-like-domain That Can Bind Poly(ADP-ribose)." J Biol Chem. 2016 Jan 8;291(2):924-38. PMID: 26565020
102. Welbourn, Sarah, and Klaus Strebel. "Low dNTP levels are necessary but may not be sufficient for lentiviral restriction by SAMHD1." Virology 488 (2016): 271-277.
103. Wolfgeher, Donald, et al. "The dynamic interactome of human Aha1 upon Y223 phosphorylation." Data in Brief 5 (2015): 752-755.
104. Carpenter, Brandon S., et al. "The heterotrimeric kinesin-2 complex interacts with and regulates GLI protein function." Journal of cell science 128.5 (2015): 1034-1050. PMID: 25588831

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Chemical Properties

Storage	Desiccate at -20°C
M.Wt	2861.87
Cas No.	402750-12-3
Formula	C₁₂₀H₁₆₉N₃₁O₄₉S
Synonyms	H-Met-Asp-Tyr-Lys-Asp-His-Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-OH
Solubility	≥25mg/ml in TBS (0.5M Tris-HCl, pH 7.4, with 1M NaCl), 1X TBS as recommended buffer.
Chemical Name	3X (DYKDDDDK) Peptide
SDF	Download SDF
Canonical SMILES	CCC(C)C(C(=O)NC(CC(=O)O)C(=O)NC(CC1=CC=C(C=C1)O)C(=O)NC(CCCCN)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CCCCN)C(=O)O)NC(=O)C(CC(=O)O)NC(=O)C(CC2=CNC=N2)NC(=O)C(CC(=O)O)NC(=O)C(CCCCN)NC(=O)C(CC3=CC=C(C=C3)O)NC(=O)C(CC(=O)O)NC(
Shipping Condition	Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips	We do not recommend long-term storage for the solution, please use it up soon.

Protocol

ELISA experiment [1]:
Preparation method	The solubility of this peptide in sterile water is >10 mM. Stock solution should be splited and stored at -80°C for several months.
Applications	3-DYKDDDDK peptide has found widespread use as a mild purification reagent for DYKDDDDK-epitope tagged recombinant proteins. Although its affinity columns release monovalent flagged proteins in the absence of calcium, the antibody retains substantial affinity for the DYKDDDDK sequence even in metal-free conditions, so that it has been impossible to use it to develop a metal-sensitive ELISA assay. This is due to the ability of the antibody to remain bound to polyvalent surface-coated antigen, for instance, when Flagged proteins are bound to ELISA plates or blotting filters. The resultant antigen polyvalence raises the avidity of the DYKDDDDK antibody to a point where the reaction is essentially calcium-independent. However, when the antibody itself was made monovalent, by proteolytic cleavage to the Fab, this situation was reversed and the ELISA reaction became calcium-dependent. This new metal-dependent ELISA assay was used to explore the metal requirements of the antibody in detail. Among divalent metals, binding tapered off with increasing radius above that of calcium, or with decreasing radius below that of calcium. Several smaller metals, such as nickel, acted as inhibitors of the binding reaction. Substantial binding was demonstrated for heavy metals such as cadmium, lanthanum and samarium. Because it is of interest to use this antibody for the co-crystallization of recombinant DYKDDDDK-fusion proteins, the ability to bind heavy metals was a significant finding.
References: 1. Hopp TP1, Gallis B, Prickett KS. Metal-binding properties of a calcium-dependent monoclonal antibody. Mol Immunol. 1996 May-Jun;33(7-8):601-8.