Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

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Anti-Tet-Repressor Antibodies

Excellent Tool for Studies of Tet Regulatory System in Eukaryotic Cells

Features

  • Suited for ELISA, Western blots and immunofluorescence (not for IHC)
  • Excellent binding properties
  • Monoclonal IgG1; κ
  • Polyclonal rabbit IgG
  • Immunogen: TetR-tetO (Accession # P04483 Pubmed)

Description

  • The tetracycline (tet) regulatory system is widely used for selective target gene regulation in eukaryotic cells. MoBiTec offers a unique set of polyclonal and monoclonal antibodies targeting the Tet-Repressor protein TetR) (TetR-tetO Accession no. P04483) for study of this popular system. These antibodies possess excellent binding properties and have been successfully tested for use in ELISA, Western blot and immunofluorescence assays.
  • Two options for the monoclonal antibodies are offered. First an optimized mix consisting of the two different epitope-specific monoclonal antibodies (TET02), and second a single monoclonal antibody, which can be used for immunofluorescence microscopy (only in cell culture) (TET03). The rabbit polyclonal antibody (TET01) can be used in all three above-mentioned applications. These antibodies provide an excellent new tool for elucidating the tet regulatory system.

Tet Repressor Antibodies are perfectly suited for detection of:

  • Tet-Repressor (TetR)
  • Tet-Repressor fusion protein (TetR-Fusion)
  • tetracycline responsive transactivator (tTA)
  • reverse tetracycline responsive transactivator (rtTA) including derivates like rtTA-S or rtTA-M
Structure of the Tet-Repressor

Structure of the Tet-
Repressor (D)-[tc•Mg]+ complex.7
The folding of the polypeptide chain is represented by a ribbon diagram. The subunits are shown in different colors. Illustration provided by Dr. E. Pook, Institute of Microbiology and Biochemistry, University of Erlangen-Nürnberg, Germany.

Note:

  • MoBiTec anti-TetR(B) antibodies were raised against TetR(B)-tetO, but are generally able to detect also variants of TetR(B), like e.g. rtTA and its derivatives, whereby TET02 is the most sensitive and promising choice. However, in contrast to TetR(B), rtTA regulator proteins are required and present in cells in much lesser amounts, especially in stable cell lines. Additionally, the regulated gene and its product may downregulate these regulators further, so that it's even more obscured. That is, detection of rtTA and its derivatives is more difficult than that of TetR(B) and often fails at the level of Western blotting. In such cases, PCR may be used as surrogate due to its sensitivity. Nevertheless, independent references, examples see below, have proven the suitability of MoBiTec's anti-TetR(B) antibodies to also detect rtTA:
    1. Markusic et al., Nucleic Acids Res., Apr 2005; 33(6): e63 (Pubmed)
    2. Benabdellah et al., PLoS One. 2011;6(8):e23734 (Pubmed)
    3. Anders et al., Transgenic Res., Oct 2012, 21(5):1099–1107 (Pubmed)
    4. arXiv:1212.5109 [q-bio.MN] (2012) (Link)

  • Therefore, MoBiTec shall not be made responsible or liable for any claims or loss arising from the failure of the anti-TetR(B) antibodies to detect rtTA and its derivatives!

Summary of the three Tet-Repressor Antibodies

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 ELISA 0.2 ng 20 - 50 pg n.d.
Detection limit Western Blot 0.8 ng 0.8 - 1.0 ng 5 ng

Examples of Different Applications

ELISA of Tet-Repressor Antibodies

ELISA of Tet-Repressor Antibodies

Immunfluorescence of Tet-Repressor Antibodies

Immunfluorescence of Tet-Repressor Antibodies

Western Blot of Tet-Repressor Antibodies

Western Blot of Tet-
Repressor Antibodies

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
TET01 anti tet-repressor, polyclonal rabbit, lyophilized 3 mg 403,00 PDF
TET02 anti tet-repressor, monoclonal IgG1, mix, lyoph. 1 mg 443,00 PDF
TET03 anti tet-repressor, monoclonal IgG1, lyoph. 50 µg 199,00 PDF
TETR1 TET Repressor protein (23 kDa), positive control 1 µg 86,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.

Literature

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
  • Cai D, Byth KF, Shapiro GI (2006): AZ703, an imidazo[1,2-a]pyridine inhibitor of cyclin-dependent kinases 1 and 2, induces E2F-1-dependent apoptosis enhanced by depletion of cyclin-dependent kinase 9. Cancer Res. 2006 Jan 1;66(1):435-44. Pubmed
  • 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., Feb 2006; 12: 1308 - 1316. 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