We believe this site might serve you best:

United States

English Continue

This country code will remain if no action is taken to change it.

Don't see your country?
Promega Corporation
Home » Resources » Tools »

Citations Search

Catalog_banner_2014

Search Within Results

Need Assistance? Chat

Sort By:

Plant J. 52, 175–184. Functional immobilization of plant receptor-like kinase onto microbeads towards receptor array construction and receptor-based ligand fishing. 2007

Shinohara, H. and Matsubayashi, Y.

Notes: The authors were looking for a method for ligand fishing experiments with plant receptor-like kinases (RLKs). The strategy chosen, functional immobilization on microbeads, used phytosulfokine (PSK) and its carrot cell receptor (DcPSKR1) as the test ligand-receptor pair. DcPSKR1 was cloned into the pHT2 HaloTag® Vector, adding the HaloTag® gene to the C terminus of DcPSKR1 or replacing the DcPSKR1 kinase domain. These constructs were transfected into BY-2 cells and expression confirmed by immunoblotting the membrane fraction and staining with DcPSKR1 and Anti-HaloTag antibodies. To confirm activity of the individual proteins, membrane fractions of BY-2 cells expressing the DcPSKR1-HaloTag® fusion proteins were tested for PSK binding activity to then HaloTag® binding activity confirmed using the HaloTag® TMR Ligand. The DcPSKR1-ΔKD-Halo protein was immobilized on HaloLink™ Resin and the Kd measured. The binding of PSK to the immobilized DcPSKR1-ΔKD-Halo protein was visualized by using fluorescently labeled Alexa488-PSK. Columns of HaloLink™ Resin with bound DcPSKR1-ΔKD-Halo protein were used to bind PSK ligands at physiological concentration, elute the ligands with a high-salt buffer and analyzed on LC-MS. (3946)

Expand Full Notes »

J. Biochem. Biophys. Methods 70, 679-682. Rapid single-tube method for small-scale affinity purification of polyclonal antibodies using HaloTagtrade Technology. 2007

Hata, T. and Nakayama, M.

Notes: This paper outlines a method for the affinity purification of polyclonal antisera using the HaloTag® Technology. The protein of interest was expressed in E. coli as a HaloTag® fusion protein. Bacterial cell lysate (100µl) was then mixed with 50µl of HaloLink™ Resin (50% suspension) for 1 hour at room temperature. After washing, 100µl of immune serum (1:10 dilution) was added and the mixture incubated for 1 hour at 4°C. The resin was then washed three times with a high-salt buffer to remove nonspecific antibodies prior to elution of specifically bound antibodies using 100mM glycine (pH 2.5). The authors discuss the advantages of this single-tube method compared to other methods of antibody purification. (3623)

Expand Full Notes »

J. Biol. Chem. 282, 13059-13072. XAF1 mediates tumor necrosis factor-α-induced apoptosis and X-linked inhibitor of apoptosis cleavage by acting through the mitochondrial pathway 2007

Straszewshi-Chavez, A., Visintin, I.P., Karassina, N., Los, G., Liston, P., Halaban, R., Fadiel, A. and Mor, G.

Notes: The authors sought to determine the mechanism by which first-trimester trophoblasts resist FAS ligand-induced apoptosis but remain sensitive to TNFα-mediated apoptosis. First trimester trophoblasts express XAF1 [X-linked inhibitor of apoptosis (XIAP)-associated factor 1], which may be involved in regulating their response to proapoptotic signals. The authors created HaloTag™-XAF1 fusion constructs and transiently transfected the first trimester trophoblast cell line (3A). Cells were labeled with the HaloTag™ TMR ligand, and XAF1 was shown to localize to the cytoplasm. 3A cells transiently transfected with the fusion construct were also separated into cytoplasmic and mitochondrial fractions. The fractions were labeled with HaloTag™ TMR ligand. Expression of the fusion peaked at 48 hours after transfection in both mitochondrial and cytoplasmic fractions. TNFα-treatment of 3A cells induced translocation of endogenous XAF1 to the mitochondria. The authors used the Caspase-Glo® Assays to demonstrate activation of caspase-3 and caspase-9 in response to expression of XAF-1. They also show that caspase-3 activation and XIAP cleavage correlate with translocation of endogenous XAF1 to mitochondria. Viability of 3A and primary trophoblasts over expressing XAF1 was evaluated using the CellTiter® 96 AQueous One-Solution Assay. (3760)

Expand Full Notes »

Angewandte Chemie International Edition 45, 4536-4940. HaloTag protein-mediated site-specific conjugation of bioluminescent proteins to quantum dots. 2006

Zhang, Y., So, M.-K., Loening, A.M., Yao, H., Gambhir, S.S., and Rao, J.

Notes: This paper describes work to use the HaloTag™ Interchangeable Protein Labeling Technology for the specific conjugation of proteins to quantum dots. The authors used the pHT2 vector to create a Renilla luciferase (Luc 8)/HaloTag™ Protein fusion that contained a 6X polyhistidine tag. The fusion protein was purified over a nickel affinity resin. They synthesized a HaloTag™ ligand for conjugation to the quantum dots. The ligand was designed to orient the ligand away from the quantum dot surface to minimize steric hindrance between the HaloTag™ fusion protein and the quantum dots upon interaction of the fusion protein with the ligand. The authors used BRET emission to evaluate conjugation of the HaloTag™ fusion protein to the quantum dots. They conclude that using this technology, they were able to conjugate a bioluminescent protein to quantum dots, creating self-illuminating quantum dots. Furthermore, they suggest that the mild conjugation conditions used may allow in vivo labeling of proteins or cells using quantum dots. (3490)

Expand Full Notes »

Products

J. Exp. Bot. July, epub ahead of print. HaloTag™: A new versatile reporter gene system in plant cells. 2006

Lang, C., Schulze, J., Mendel, R-R. and Hänsch, R.

Notes: This paper highlights the first use of the HaloTag™ Interchangeable Protein Labeling Technology in plant cells. The cDNA for the HaloTag™ protein was amplified by PCR from the pHT2 Vector and cloned into the pGEM®-T Easy® Vector, from which it was excised and transferred to a second vector where its expression was under the control of the cauliflower mosaic virus (CaMV)-35S promoter. The construct was transformed into tobacco protoplasts and bioballistically transformed into tobacco leaf cells. Localization was followed using the HaloTag™ TMR and diAcFAM Ligands. (3503)

Expand Full Notes »

Cell 126, 335-348. Single-molecule analysis of dynein processivity and stepping behavior. 2006

Reck-Peterson, S.L., Yidiz, A., Carter, A.P., Gennerich, A., Zhang, N. and Vale, R.D.

Notes: HaloTag™ Interchangeable Labeling Technology was used to specifically label engineered dynein produced in Saccharomyces cerevisiae. The HaloTag™ Protein was added in-frame with either the 5´ or 3´ end of the coding sequence of various engineered dynein molecules. The authors report being able to label the dynein in specific locations using fluorescent dyes or quantum dots. HaloTag™ TMR Ligand was used to covalently label dynein to directly visualize dynein motor movement using total internal reflection fluorescence microscopy. HaloTag™ Biotin Ligand was used to label dynein with streptavidin quantum dots. (3504)

Expand Full Notes »

It appears that you have Javascript disabled. Our website requires Javascript to function correctly. For the best browsing experience, please enable Javascript.

Scientists at Your Service

Scientists at Your Service

We offer a range of services to help you succeed using Promega technologies. From product training to set up of automated systems and development of custom applications—our scientific support goes beyond the basics.

Ask us! We are here to help you.