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| 1. |
Hurst, R., Hook, B., Slater, M.R., Hatrnett, J., Storts, D.R., and Nath, N.
(2009)
Protein-protein interaction studies on protein arrays: effect of detection strategies on signal-to-background ratios.
Anal Biochem.
392
,
45-53
.
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Notes:
These authors compared 6 different detection strategies for protein-protein interactions on protein arrays. They expressed HaloTag® labeled bait proteins in a cell-free expression system, and captured these bait proteins onto coated glass slides using the HaloLink™ Array System. They then compared detection strategies using prey proteins labeled as follows: 1)35S methionine, 2) fluorescence (BODIPY-FL) and 3) biotin labeling of lysine residues using modified Lys tRNA, 4) chemical labeling after expression, 5) HaloTag® fusion, and 6) N-terminal FLAG tag. The authors evaluated signal:background ratios, adaptability to high-throughput screening, and ease of use.
(0003999) |
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Products: FluoroTect™ GreenLys in vitro Translation Labeling System | HaloLink™ Array (TNT® SP6 Wheat Germ) Two Slide System | HaloTag® Biotin Ligand | HaloTag® TMR Ligand | pF1A T7 Flexi® Vector | pFN19K HaloTag® T7 SP6 Flexi® Vector | TNT® SP6 Quick Coupled Transcription/Translation System, Trial Size | Transcend™ tRNA |
| 2. |
Shibuya, N., and Nakashima, N.
(2008)
Characterization of the 5' internal ribosome entry site of Plautia stali intestine virus.
J. Gen. Virol.
87
,
3679-3686
.
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Notes:
The Plautia stali virus contains two open reading frames and includes a 5´ internal ribosome entry site (IRES) and an intergenic IRES region. These authors showed that the 5´ IRES was functional and initiated translation in insect cell lysate, but not in rabbit reticulocyte lysate or wheat germ extract. The efficiency of translation mediated by the 5´ IRES region was tested with and without cap analogue using various firefly and Renilla luciferase reporter constructs. They also used deletion mutants to identify the specific regions required for translation initiation.
(0003942) |
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Products: FluoroTect™ GreenLys in vitro Translation Labeling System | pSP-luc+NF Fusion Vector | Transcend™ tRNA |
| 3. |
He, W., Shi, Q., Hu, X. and Yan, R.
(2007)
The membrane topology of RTN3 and its effect on binding of RTN3 to BACE1
J. Biol. Chem.
282
,
29144-29151
.
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Notes:
The authors of this study determined the membrane topology of reticulon 3 (RTN3), an integral membrane protein that is expressed at high levels in neruons and has been show to negatively regulate the activity of BACE1 (Beta site APP-Cleaving Enzyme). Disruption of RTN3 is associated with incidence of dystrophic neurites in AD brain. RTN3 was translated using the TNT® Quick Coupled Transcription/Translation System in the presence of Canine Microsomal Membranes and labeled using the Transcend™ Non-Radioactive Translation Detection System.
(0003860) |
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Products: Canine Pancreatic Microsomal Membranes | TNT® SP6 Quick Coupled Transcription/Translation System | TNT® SP6 Quick Coupled Transcription/Translation System, Trial Size | TNT® T7 Quick Coupled Transcription/Translation System | TNT® T7 Quick Coupled Transcription/Translation System, Trial Size | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System |
| 4. |
Baldwin, A., Huh, K-W. and Mϋnger, K.
(2006)
Human papillomavirus e7 oncoprotein dysregulates steroid receptor coactivator 1 localization and function.
J. Virol.
80
,
6669-6677
.
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Notes:
The MagneGST™ Protein Purification System was used to purify GST fusion proteins of the oncoprotein HPV16 E7 or various mutants of HPV16 E7. The purified GST fusion proteins were used for in vitro binding experiments with steroid receptor coactivator 1 (SRC-1), which was produced using the TNT® T7 Coupled Wheat Germ Extract System and labeled with the Transcend™ Non-Radioactive Translation Detection System. GST pull-down assays were resolved by Western analysis using streptavidn-horseradish peroxidase and alpha-GST. To determine the effects of endogenously expressed HPV16 E7 on SCR-1-mediated transcription, luciferase reporters under the control of either the IL-8 promoter or an artificial promoter containing three estrogen response elements repeats (3 × EREs) were cotransfected with a Renilla control vector into two human cervical cancer lines (C33A and CaSki) using either the Transfast™ Transfection Reagent or another commercial transfection reagent. The Dual-Luciferase® Reporter Assay was then used to determine luciferase activity to functionally map the E7-interacting domain and to determine the effects of high- and low-risk PHV E7s on SRC-1-mediated transcription.
(0003459) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | MagneGST™ Glutathione Particles | MagneGST™ Protein Purification System | TNT® T7 Coupled Wheat Germ Extract System | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System | Transcend™ tRNA | TransFast™ Transfection Reagent |
| 5. |
Zheng Q., Yin G., Yan C., Cavet M. and Berk B.C.
(2004)
14-3-3beta binds to big mitogen-activated protein kinase 1 (BMK1/ERK5) and regulates BMK1 function.
J. Biol. Chem.
279(10)
,
8787-8791
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Notes:
The authors performed a yeast two-hybrid screen using big mitogen-activated kinase 1 (BMK1/ERK5) as the bait and identified the scaffolding protein 14-3-3beta. To confirm this interaction, the cloned mouse BMK1 gene was expressed in the TNT® T7 Quick Coupled Transcription/Translation System. The expressed protein was labeled with Transcend™ tRNA. Using a GST-14-3-3beta fusion protein, a pull-down assay was performed and the direct binding confirmed after immunoblotting and staining with streptavidin-horseradish peroxidase (HRP). The interaction of various BMK1 mutants were tested in a mammalian two-hybrid system and measured by the Dual-Luciferase® Reporter Assay System.
(0003078) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | TNT® T7 Quick Coupled Transcription/Translation System | TNT® T7 Quick Coupled Transcription/Translation System, Trial Size | Transcend™ tRNA |
| 6. |
Weinhofer, I., Forss-Petter, S., Zigman, M. and Berger, J.
(2002)
Cholesterol regulates ABCD2 expression: Implications for the therapy of X-linked adrenoleukodystrophy.
Hum. Mol. Genet.
11
,
2701–2708
.
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Notes:
The sterol regulatory element-binding protein-1a (SREBP1a) was in vitro transcribed and translated from a plasmid template using TNT® T7 Quick for PCR DNA. A control reaction was performed using Transcend™ tRNA to confirm that the 120KDa protein was expressed correctly. Unlabeled SREBP1a was used in gel-shift assays with a labeled oligo representing the SRE motif from the human ABCD2 gene promoter.
(0003222) |
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Products: TNT® T7 Quick for PCR DNA | Transcend™ tRNA |
| 7. |
van der Luijt, R.B., van Zon, P.H., Jansen, R.P., van der Sijs-Bos, C.J., Warlam-Rodenhuis, C.C. and Ausems, M.G.
(2001)
De novo recurrent germline mutation of the BRCA2 gene in a patient with early onset breast cancer.
J. Med. Genet.
38
,
102-105
.
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Notes:
This paper describes using the TNT® T7 Coupled Reticulocyte Lysate System and Transcend™ tRNA to perform a nonradioactive in vitro Protein Truncation Test for a BRCA2 gene product. Templates for the transcription-translation reactions were created by PCR-amplifying regions of the BRCA2 gene from normal and cancer patient sample genomic DNA.
(0003075) |
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Products: TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System | Transcend™ tRNA |
| 8. |
Pei, L.
(2000)
Phosphorylation modulates the function of the vasoactive intestinal polypeptide receptor transcriptional repressor protein.
J. Biol. Chem.
275
,
1176-1182
.
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Notes:
The TNT® Coupled Reticulocyte Lysate System was used in conjunction with the Transcend™ Chemiluminescent Non-Radioactive Translation Detection System. Various truncations of the protein of interest were produced ranging from 67kDa to 19kDa to use in gel shift assays. The truncations were used to indentify which region bound the oligo of interest. The in vitro translated proteins were also used with antibodies for supershifts.
(0000003) |
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Products: TNT® SP6 Coupled Reticulocyte Lysate System | TNT® SP6 Coupled Reticulocyte Lysate System, Trial Size | TNT® SP6 Quick Coupled Transcription/Translation System | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | TNT® T7 Quick Coupled Transcription/Translation System | TNT® T7/SP6 Coupled Reticulocyte Lysate System | TNT® T7/T3 Coupled Reticulocyte Lysate System | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System | Transcend™ tRNA |
| 9. |
Chatterjee-Kishore, M. , van Den Akker, F. , and Stark, G. R.
(2000)
Adenovirus E1A down-regulates LMP2 transcription by interfering with the binding of stat1 to IRF1.
J. Biol. Chem.
275(27)
,
20406-20411
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Notes:
Various Stat-1 and IRF-1 proteins were expressed in vitro with the TNT® T7 Coupled Reticulocyte Lysate System for various protein-protein interaction studies by co-immunoprecipitation with antibodies followed by immunoblotting. To insure equal amounts of proteins were expressed and intact, the Transcend™ Biotinylated tRNA was incorporated during the reaction and a portion of the reaction was removed, electrophoresed, transfered and detected with an HRP-conjugate.
(0000030) |
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Products: TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | Transcend™ tRNA |
| 10. |
Bazzoni, G. , Martinez Estrada, O. M. , Orsenigo, F. , Cordenonsi, M. , Citi, S. , and Dejana, E.
(2000)
Interaction of junctional adhesion molecule with the tight junction components ZO-1, cingulin, and occludin.
J. Biol. Chem.
275(27)
,
20520-20526
.
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Notes:
The 1736 amino acid (predicted MW of 194kDa) protein ZO-1 was translated in vitro with the TNT® T7 Coupled Reticulocyte System and the Transcend™ Biotinylated tRNA. There were 107 lysines in the protein. The protein was used in protein-protein interaction studies. A protein, JAM, was expressed as a GST fusion in E. coli and immobilized to a glutathione-linked sepharose beadsor peptides were immobilized to sepharose beads. The translation extract was interacted with the beads, washed and bound protein removed by boiling in SDS sample buffer. The bound protein was electrophoresed, transfered and detected via the biotin tag on the ZO-1 protein.
(0000031) |
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Products: TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | Transcend™ tRNA |
| 11. |
Chien, W. , and Pei, L.
(2000)
A novel binding factor facilitates nuclear translocation and transcriptional activation function of the pituitary tumor-transforming gene product.
J. Biol. Chem.
275(25)
,
19422-19427
.
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Notes:
The TNT® T3 Coupled Reticulocyte Lysate System was used to express the PTTG-binding factor, a 179 amino acid protein (predicted 19.7kDa). The protein was translated in the presence of the Transcend™ Biotinylated tRNA. The expressed protein was used to GST-fusion protein pulldown reactions with a PTTG-GST fusion. The bound proteins were removed by boiling in SDS sample buffer, electrophoresed, transfered, and detected by chemiluminescent HRP.
(0000032) |
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Products: TNT® T3 Coupled Reticulocyte Lysate System | Transcend™ tRNA |
| 12. |
Morgan, H. , Smith, M. , Burke, Z. , and Carter, D.
(2000)
The transactivation-competent carboxyl-terminal domain of AF-9 is expressed within a sexually dimorphic transcript in rat pituitary.
FASEB J.
14(9)
,
1109-1116
.
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Notes:
A clone of the rat AF-9 cDNA was isolated. Two potential start codons were present in the cDNA. To see which is prefered by a mammalian translation system, the cDNA was translated in vitro with the TNT® Coupled Reticulocyte Lysate System and the Transcend™ Biotinylated tRNA. The preferred transcript was the first ATG resulting in the 370 amino acid (40.7kDa predicted molecular weight) which had 47 lysines.
(0000033) |
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Products: TNT® SP6 Coupled Reticulocyte Lysate System | TNT® SP6 Coupled Reticulocyte Lysate System, Trial Size | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | Transcend™ tRNA |
| 13. |
Yamamoto, K., Masuno, H., Choi, M., Nakashima, K., Taga, T., Ooizumi, H., Umesono, K., Sicinska, W., VanHooke, J., DeLuca, H.F., Yamada, S.
(2000)
Three-dimensional modeling of and ligand docking to vitamin D receptor ligand binding domain.
Proc. Natl. Acad. Sci. U S A
97
,
1467-1472
.
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Notes:
The TNT® Coupled Reticulocyte Lysate System and the Transcend™ Chemiluminescent Non-Radioactive Translation Detection System were used to produce and label wildtype and mutant human vitamin D receptors. The proteins produced were used for vitamin D binding assays.
(0000133) |
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Products: TNT® SP6 Coupled Reticulocyte Lysate System | TNT® SP6 Coupled Reticulocyte Lysate System, Trial Size | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | TNT® T7/SP6 Coupled Reticulocyte Lysate System | TNT® T7/T3 Coupled Reticulocyte Lysate System | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System |
| 14. |
Wen, F. , Zhu, Y. , and Hawes, M. C.
(1999)
Effect of pectin methylesterase gene expression on pea root development.
Plant Cell
11(6)
,
1129-1140
.
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Notes:
The cDNA of the pectin methyltransferase was translated in vitro with the TNT® Coupled Reticulocyte Lysate System in the presence of the Transcend™ Biotinylated tRNA. The protein was used for in vitro enzymatic assay of activity. The assay was colorimetric and the heme in the rabbit lysate interferred. The hemoglobin was removed by acid precipitation and the method is referenced. The enzyme contains 555 amino acids and 39 lysines.
(0000029) |
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Products: TNT® SP6 Coupled Reticulocyte Lysate System | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | Transcend™ tRNA |
| 15. |
Pei, L.
(1999)
Pituitary tumor-transforming gene protein associates with ribosomal protein S10 and a novel human homologue of DnaJ in testicular cells.
J. Biol. Chem.
274
,
3151-3158
.
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Notes:
The authors used the TNT® Coupled Reticulocyte Lysate System with Transcend™ Biotinylated tRNA to synthesize S10 and HSJ2 protein for protein:protein interaction studies with GST-tagged PTTG.
(0000556) |
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Products: TNT® SP6 Coupled Reticulocyte Lysate System | TNT® SP6 Coupled Reticulocyte Lysate System, Trial Size | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | TNT® T7/SP6 Coupled Reticulocyte Lysate System | TNT® T7/T3 Coupled Reticulocyte Lysate System | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System | Transcend™ tRNA |
| 16. |
Hussain, N.K., Yamabhai, M., Ramjaun, A.R., Guy, A.M., Baranes, D., O'Bryan, J.P., Der, C.J., Kay, B.K., McPherson, P.S.
(1999)
Splice variants of intersectin are components of the endocytic machinery in neurons and nonneuronal cells.
J. Biol. Chem.
274
,
15671-15677
.
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Notes:
The proteins Ibp2, MP90 and luciferase were translated in vitro with the TNT® Coupled Reticulocyte Lysate System in the presence of the Transcend™ Biotinylated tRNA. The translated proteins were reacted with a glutathione bead-immobilized portion of the clathrin heavy chain. Bound proteins were solubilized in SDS sample buffer and the proteins detected by Western detection with the Transcend™ Non-Radioactive Translation Detection System.
(0000990) |
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Products: TNT® SP6 Coupled Reticulocyte Lysate System | TNT® SP6 Coupled Reticulocyte Lysate System, Trial Size | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | TNT® T7/SP6 Coupled Reticulocyte Lysate System | TNT® T7/T3 Coupled Reticulocyte Lysate System | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System | Transcend™ tRNA |
| 17. |
Estelles, A. , Yokoyama, M. , Nothias, F. , Vincent, J. D. , Glowinski, J. , Vernier, P. , Chneiweiss, H.
(1996)
The major astrocytic phosphoprotein PEA-15 is encoded by two mRNAs conserved on their full length in mouse and human.
J. Biol. Chem.
271
,
14800-14806
.
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Notes:
PEA-15 cDNAs were cloned from a mouse astrocytic library, and the clones were linearized and expressed using T3 or T7 Polymerase in the TNT® Reticulocyte Lysate System either in the presence of [35S]methionine or Transcend™ tRNA. The resulting proteins were analyzed by SDS-PAGE and Western blotting, respectively. A unique 15kDa protein that was recognized by an antibody to PEA-15 was produced in the TNT® System.
(0001178) |
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Products: TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size | TNT® T7/T3 Coupled Reticulocyte Lysate System | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System | Transcend™ tRNA |
| 18. |
Sanford, J.C., Yu, J., Pan, J.Y., Wessling Resnick, M.
(1995)
GDP dissociation inhibitor serves as a cytosolic acceptor for newly synthesized and prenylated Rab5.
J. Biol. Chem.
270
,
26904-26909
.
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Notes:
In this work, Sanford and colleagues study the GTP-binding protein, Rab5, by expressing this protein in Rabbit Reticulocyte Lysate (RRL) with Transcend™ Biotinylated tRNA. In RRL, prenylated biotin-Rab5 associates with a 45kDa GDP dissociation inhibitor in a ~70kDa particle on sucrose density gradients. The authors suggest that biotin-Rab5 provides a novel tool for capturing and characterizing accessory factors required for Rab protein function.
(0000436) |
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Products: Rabbit Reticulocyte Lysate System, Nuclease Treated | Transcend™ tRNA |