We are pleased to offer a set of polyclonal and monoclonal anti-Tet-Repressor (TetR) antibodies, manufactured by MoBiTec. The immunogen is TetR-tetO (Accession no. P04483 Pubmed). These antibodies possess excellent binding properties and have been successfully tested for use in ELISA, Western blot and immunofluorescence assays. They can be used in the detection of Tet-Repressor (TetR), Tet-Repressor fusion protein (TetR-Fusion), tetracycline responsive transactivator (tTA) and reverse tetracycline responsive transactivator (rtTA). We also offer a TET-Repressor Protein as a positive control, see item# TETR1.

TET01 is a polyclonal rabbit anti-Tet-Repressor antibody. It was prepared using TetR-tetO as immunogen, and has been purified on Protein G. It is provided as 200ul of purified antibody, lyophilized. After reconstitution to 200µl in distilled water, it can be further diluted 1:1000 for Western blots and ELISA applications. (Monoclonal antibody TET03 is recommended for immunofluorescence studies.) This antibody has excellent binding properties to TetR. Detection limits with this antibody are as follows: ELISA = 0.2ng; Western Blots = 0.8ng.

TET02 is provided as 1mg of purified antibody, lyophilized. After reconstitution to 100ul in distilled water, it can be further diluted 1:500 - 1:2000 for Western blots and ELISA applications. (Monoclonal antibody TET03 is recommended for immunofluorescence studies.) This antibody has excellent binding properties to TetR. Detection limits with this antibody are as follows: ELISA = 20-50pg; Western Blots = 0.8 - 1ng

TET03 is a mouse monoclonal antibody, IgG1 Kappa. The antibody recognizes amino acid # 37-44 and is ideal for immunofluorescence studies. TET03 is provided as 50µg of purified antibody, lyophilized. After reconstitution to 100µl in distilled water, it can be further diluted 1:100 to 1:500 for immunofluorescence studies.

	TET01	TET02	TET03
Type	Rabbit polyclonal IgG	monoclonal IgG1; κ mix	monoclonal IgG1; κ
Immunogen	TetR-tetO	TetR-tetO	TetR-tetO
Purification	Affinity purified via Protein G columns	Affinity purified via Protein A or G columns	Affinity purified via Protein A or G columns
Epitope	--	TetR: -Amino acid #84-98 -Amino acid #26-53	TetR: -Amino acid #37-44
Reconstitution In:	200 µl dest. H₂O	100 µl dest. H₂O	100 µl dest. H₂O
Working dilution for immunofluorescence	n.d.	n.d.	1:100 - 1:500
Working dilution for Western Blots and ELISA	1:1000	1:500 - 1:2000	1:1000
Detection limit	0.2 ng	20 - 50 pg	n.d.
Detection limit for Western Blot	0.8 ng	0.8 - 1.0 ng	5 ng

Features

Prime tool for studying tet regulatory systems in eukaryotic cells
Suited for ELISA, Western blots and Immunofluorescence (for cells only, not for tissues or tissue slices)
Excellent binding properties

Applications

Detection of Tet-Repressor (TetR)
Detection of Tet-Repressor fusion protein (TetR-Fusion)
Detection of tetracycline responsive transactivator (tTA)
Detection of reverse tetracycline responsive transactivator (rtTA) such as rtTA-S or rtTA-M

TET01 - anti tet-repressor, polyclonal rabbit, lyophilized, 3mg
TET02 - anti tet-repressor, monoclonal IgG1, mix, lyophilized, 1mg
TET03 - anti tet-repressor, monoclonal IgG1, lyophilized, 50 µg

Literature

Product Information Sheet

Publications

Tet Regulatory System

Pook E, Grimm S, Bonin A, Winkler T, Hillen W.(1998): Affinities of mAbs to Tet repressor complexed with operator or tetracycline suggest conformational changes associated with induction. Eur J Biochem. 1998 Dec 15;258(3):915-22. Pubmed

Hillen W, Berens C.(1994): Mechanisms underlying expression of Tn10 encoded tetracycline resistance. Annu Rev Microbiol. 1994;48:345-69. Pubmed

Gatz C.(1995): Novel inducible/repressible gene expression systems. Methods Cell Biol. 1995;50:411-24. Pubmed

Mayford M, Bach ME, Huang YY, Wang L, Hawkins RD, Kandel ER.(1996): Control of memory formation through regulated expression of a CaMKII transgene. Science. 1996 Dec 6;274(5293):1678-83. Pubmed

Hwang JJ, Scuric Z, Anderson WF. (1996): Novel retroviral vector transferring a suicide gene and a selectable marker gene with enhanced gene expression by using a tetracycline-responsive expression system. J Virol. 1996 Nov;70(11):8138-41. Pubmed Free

Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H. (1996): Transcriptional activation by tetracyclines in mammalian cells. Science. 1995 Jun 23;268(5218):1766-9. Pubmed

Hinrichs W, Kisker C, Duvel M, Muller A, Tovar K, Hillen W, Saenger W (1994): Structure of the Tet repressor-tetracycline complex and regulation of antibiotic resistance. Science. 1994 Apr 15;264(5157):418-20. Pubmed

References with Tet-Repressor (TetR) Antibodies

Marti F, Xu CW, Selvakumar A, Brent R, Dupont B, King PD. (1998): LCK-phosphorylated human killer cell-inhibitory receptors recruit and activate phosphatidylinositol 3-kinase. Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11810-5. Pubmed Free

Freundlieb S, Schirra-Muller C, Bujard H.(1999): A tetracycline controlled activation/repression system with increased potential for gene transfer into mammalian cells. J Gene Med. 1999 Jan-Feb;1(1):4-12. Pubmed

Urlinger S, Baron U, Thellmann M, Hasan MT, Bujard H, Hillen W. (2000): Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):7963-8. Pubmed Free

Meissner M, Brecht S, Bujard H, Soldati D. (2001): Modulation of myosin A expression by a newly established tetracycline repressor-based inducible system in Toxoplasma gondii. Nucleic Acids Res. 2001 Nov 15;29(22):E115. Pubmed Free

Lorenz P, Koczan D, Thiesen HJ (2001): Transcriptional repression mediated by the KRAB domain of the human C₂H₂ zinc finger protein Kox1/ZNF10 does not require histone deacetylation. Biol Chem. 2001 Apr;382(4):637-44. Pubmed

Fiedler M, Skerra A (2001): proBA complementation of an auxotrophic E. coli strain improves plasmid stability and expression yield during fermenter production of a recombinant antibody fragment. Gene. 2001 Aug 22;274(1-2):111-8. Pubmed

Stebbins MJ, Urlinger S, Byrne G, Bello B, Hillen W, Yin JC (2001): Tetracycline-inducible systems for Drosophila. Proc Natl Acad Sci U S A. 2001 Sep 11;98(19):10775-80. Epub 2001 Aug 21. Pubmed Free

David KM, Perrot-Rechenmann C (2001): Characterization of a tobacco Bright Yellow 2 cell line expressing the tetracycline repressor at a high level for strict regulation of transgene expression. Plant Physiol. 2001 Apr;125(4):1548-53. Pubmed Free

Reeves PJ, Callewaert N, Contreras R, Khorana HG (2002): Structure and function in rhodopsin: high-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line. Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13419-24. Epub 2002 Oct 7. Pubmed Free

Kamper MR, Gohla G, Schluter G (2002): A novel positive tetracycline-dependent transactivator (rtTA) variant with reduced background activity and enhanced activation potential. FEBS Lett. 2002 Apr 24;517(1-3):115-20. Pubmed

Krueger C, Berens C, Schmidt A, Schnappinger D, Hillen W (2003): Single-chain Tet transregulators. Nucleic Acids Res. 2003 Jun 15;31(12):3050-6. Pubmed Free

Czauderna F, Santel A, Hinz M, Fechtner M, Durieux B, Fisch G, Leenders F, Arnold W, Giese K, Klippel A, Kaufmann J (2003): Inducible shRNA expression for application in a prostate cancer mouse model. Nucleic Acids Res. 2003 Nov 1;31(21):e127. Pubmed Free

Leenders F, Mopert K, Schmiedeknecht A, Santel A, Czauderna F, Aleku M, Penschuck S, Dames S, Sternberger M, Rohl T, Wellmann A, Arnold W, Giese K, Kaufmann J, Klippel A (2004): PKN3 is required for malignant prostate cell growth downstream of activated PI 3-kinase. EMBO J. 2004 Aug 18;23(16):3303-13. Epub 2004 Jul 29. Pubmed Free

Das AT, Zhou X, Vink M, Klaver B, Verhoef K, Marzio G, Berkhout B (2004): Viral evolution as a tool to improve the tetracycline-regulated gene expression system. J Biol Chem. 2004 Apr 30;279(18):18776-82. Epub 2004 Feb 2. Pubmed Free

Rupp B, Ruzsics Z, Sacher T, Koszinowski UH (2005): Conditional cytomegalovirus replication in vitro and in vivo. J Virol. 2005 Jan;79(1):486-94. Pubmed Free

Markusic D, Oude-Elferink R, Das AT, Berkhout B, Seppen J (2005): Comparison of single regulated lentiviral vectors with rtTA expression driven by an autoregulatory loop or a constitutive promoter. Nucleic Acids Res. 2005 Apr 4;33(6):e63. Pubmed Free

Osada M, Park HL, Nagakawa Y, Yamashita K, Fomenkov A, Kim MS, Wu G, Nomoto S, Trink B, Sidransky D (2005): Differential recognition of response elements determines target gene specificity for p53 and p63. Mol Cell Biol. 2005 Jul;25(14):6077-89. Pubmed Free

Ito T, Hashimoto Y, Tanaka E, Kan T, Tsunoda S, Sato F, Higashiyama M, Okumura T, Shimada Y (2006): An inducible short-hairpin RNA vector against osteopontin reduces metastatic potential of human esophageal squamous cell carcinoma in vitro and in vivo. Clin Cancer Res. 2006 Feb 15;12(4):1308-16. Pubmed

Lukes J, Paris Z, Regmi S, Breitling R, Mureev S, Kushnir S, Pyatkov K, Jirku M, Alexandrov KA (2006): Translational initiation in Leishmania tarentolae and Phytomonas serpens (Kinetoplastida) is strongly influenced by pre-ATG triplet and its 5' sequence context. Mol Biochem Parasitol. 2006 Aug;148(2):125-32. Epub 2006 Apr 18. Pubmed

Cai D, Vaughan M. Latham, Jr., Xinxin Zhang, and Geoffrey I. Shapiro (2006): Combined Depletion of Cell Cycle and Transcriptional Cyclin-Dependent Kinase Activities Induces Apoptosis in Cancer Cells. Cancer Res., Sep 2006; 66: 9270 - 9280. Pubmed

Bulliard Y, Wiznerowicz M, Barde I, Trono D (2006): KRAB Can Repress Lentivirus Proviral Transcription Independently of Integration Site J. Biol. Chem., Nov 2006; 281: 35742 - 35746. Pubmed

Palona I, Namba H, Mitsutake N, Starenki D, Podtcheko A, Sedliarou I, Ohtsuru A, Saenko V, Nagayama Y, Umezawa K, Yamashita S (2006): BRAFV600E promotes invasiveness of thyroid cancer cells through nuclear factor kappaB activation. Endocrinology, Dec 2006; 147: 5699 - 5707. Pubmed

Seibler J, Kleinridders A, Küter-Luks B, Niehaves S, Brüning JC, Schwenk F (2007): Reversible gene knockdown in mice using a tight, inducible shRNA expression system. Nucleic Acids Res., Apr 2007; 35: e54. Pubmed

Carrillo J, García-Aragoncillo E, Azorín D, Agra N, Sastre A, González-Mediero I, García-Miguel P, Pestaña A, Gallego S, Segura D, Alonso J (2007): Cholecystokinin Down-Regulation by RNA Interference Impairs Ewing Tumor Growth Clin. Cancer Res., Apr 2007; 13: 2429 - 2440. Pubmed

Wegmüller D, Raineri I, Gross B, Oakeley EJ, Moroni C (2007): A Cassette System to Study Embryonic Stem Cell Differentiation by Inducible RNA Interference. Stem Cells, May 2007; 25: 1178 - 1185. Pubmed

Weber A, Paschen SA, Heger K, Wilfling F, Frankenberg T, Bauerschmitt H, Seiffert BM, Kirschnek S, Wagner H, Häcker G (2007): BimS-induced apoptosis requires mitochondrial localization but not interaction with anti-apoptotic Bcl-2 proteins. J. Cell Biol., May 2007; 177: 625 - 636. Pubmed

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Anti-Tet-Repressor Antibodies

TET Ab - Anti-Tet-Repressor Antibodies

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Literature

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