Generation of dual-variable-domain immunoglobulin molecules for dual-specific targeting.

abstract：:White adipose tissue (WAT) is a heterogeneous tissue composed of lipid-filled adipocytes and several nonadipocyte cell populations, including endothelial, blood, uncharacterized stromal, and adipocyte precursor cells. Although lipid-filled adipocytes account for the majority of WAT volume and mass, nonadipocyte cell p...

journal_title：Methods in enzymology

authors： Church CD,Berry R,Rodeheffer MS

更新日期：2014-01-01 00:00:00

abstract：:The procedures described in this chapter have enabled us to identify and characterize monoclonal antibodies and their respective anti-idiotypes. We have developed several different types of immunoassays which afford greater flexibility to the investigator, depending on the type of antibodies desired and the availabili...

journal_title：Methods in enzymology

authors： Kussie PH,Albright G,Linthicum DS

更新日期：1989-01-01 00:00:00

abstract：:Death receptors are a subgroup of the tumor necrosis factor receptor superfamily (TNFRSF) and mediate activation of what is widely known as the "extrinsic" apoptosis pathway. TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is one of the most recent death receptor ligands identified. The TRAIL receptor ...

journal_title：Methods in enzymology

authors： Harper N,MacFarlane M

更新日期：2008-01-01 00:00:00

abstract：:RNA modifications, with over one hundred known so far, are commonly proposed to fine-tune the structure and function of RNA. While modifications in rRNA and tRNA are used to modulate RNA folding and decoding properties, little is known about the function of internal modifications in mRNA/lncRNA, which includes N(6)-me...

journal_title：Methods in enzymology

authors： Liu N,Pan T

更新日期：2015-01-01 00:00:00

abstract：:Organ-specific delivery of biofunctional agents is thought to enhance their activity and to reduce their side effects. Liposomes have been used as drug carriers in cancer chemotherapy, since they accumulate passively in tumor tissues due to an enhanced permeability and retention (EPR) effect. In addition, modification...

journal_title：Methods in enzymology

authors： Asai T,Oku N

更新日期：2005-01-01 00:00:00

abstract：:G-protein-coupled receptors (GPCRs) represent a major class of receptors through which a number of signals ranging from photons to large glycoprotein hormones are recognized. Human genome encodes about 800 GPCRs, yet very little structural information is available on this class of receptors. Structural studies provide...

journal_title：Methods in enzymology

authors： Ashok Y,Nanekar RT,Jaakola VP

更新日期：2013-01-01 00:00:00

abstract：:Natural products continue to demonstrate their utility both as therapeutics and as molecular probes for the discovery and mechanistic deconvolution of various cellular processes. However, this utility is dampened by the inherent difficulties involved in isolating and characterizing new bioactive natural products, in o...

journal_title：Methods in enzymology

authors： Low WK,Dang Y,Schneider-Poetsch T,Shi Z,Choi NS,Rzasa RM,Shea HA,Li S,Park K,Ma G,Romo D,Liu JO

更新日期：2007-01-01 00:00:00

abstract：:Survival of macrophages, which serve as the first-line defense against invading pathogens by invoking the overproduction of highly toxic peroxynitrite (ONOO-), depends on their ability to maintain the intracellular GSH level and to induce the expression of heme oxygenase-1 (HO-1). The ONOO- is produced by macrophages ...

journal_title：Methods in enzymology

authors： Srisook K,Kim C,Cha YN

更新日期：2005-01-01 00:00:00

abstract：:Two lymphoid cell-specific proteins, called RAG-1 and RAG-2, initiate the process of antigen receptor gene rearrangement, termed V(D)J recombination, by assembling a protein-DNA complex with two recombination signal sequences (RSSs), each of which adjoins a different receptor gene segment, and then introducing a DNA d...

journal_title：Methods in enzymology

authors： Bergeron S,Anderson DK,Swanson PC

更新日期：2006-01-01 00:00:00

abstract：:The three major considerations in reducing metal-gel binding are (1) the type of gel to be used, (2) the ionic strength of the eluent, and (3) alkaline reduction of the gel. Each of these parameters should be considered in advance of an experiment. A method is offered for determining the optimal gel type, gel treatmen...

journal_title：Methods in enzymology

authors： Martin MT

更新日期：1988-01-01 00:00:00

abstract：:Pulsed EPR methods for the study of drug binding to heme-thiolate enzymes such as cytochrome P450 and nitric oxide synthase are discussed. HYSCORE and ENDOR methods to measure (1)H of axial ligands of the heme group are described with illustrations of water serving as the axial ligand in the drug-free enzyme and ligan...

journal_title：Methods in enzymology

authors： Cruce AA,Lockart M,Bowman MK

更新日期：2015-01-01 00:00:00

abstract：:The yeast Saccharomyces cerevisiae is an ideal model system for examining fundamental nitrogen oxide biochemistry. The utility of this model system lies in both the similarities and the differences between yeast and mammalian cells. The similarities between the two systems, with regards to many of the fundamental bioc...

journal_title：Methods in enzymology

authors： Shinyashiki M,Lopez BE,Rodriguez CE,Fukuto JM

更新日期：2005-01-01 00:00:00

abstract：:The innate immune system provides host defense against a variety of threats, including microbial infection and cellular damage. Pattern recognition receptors identify both pathogen and damage associated molecular motifs associated with these events. Nucleic acid is one such danger signal that is detected by a set of c...

journal_title：Methods in enzymology

authors： Antiochos B

更新日期：2019-01-01 00:00:00

abstract：:The analysis of a reaction pathway by stopped-flow and chemical quench-flow methods provides the only means of directly measuring the rates of reaction at the active site of an enzyme. The interpretation of data from each technique is similar and involves solution of the differential equations describing the approach ...

journal_title：Methods in enzymology

authors： Johnson KA

更新日期：1986-01-01 00:00:00

abstract：:Phosphoinositide-3-kinase (PI3K) signaling has been implicated in a panoply of cellular responses including survival, proliferation, protein synthesis, migration, and vesicular trafficking. In addition, alterations in the enzymatic activity of PI3Ks have been involved in the pathogenesis of multiple diseases, ranging ...

journal_title：Methods in enzymology

authors： Ciraolo E,Gulluni F,Hirsch E

更新日期：2014-01-01 00:00:00

abstract：:Ubiquitination is a prevalent protein modification catalyzed by E1, E2, and E3 enzymes that activate, conjugate, and ligate, respectively, the ubiquitin protein to substrate protein. In order to establish a mutualistic or parasitic relationship with their eukaryotic hosts, many microorganisms hijack different aspects ...

journal_title：Methods in enzymology

authors： Qiu J,Luo ZQ

更新日期：2019-01-01 00:00:00

abstract：:The combination of PCR amplification and chemiluminescent detection of PCR products provides a highly sensitive system for the quantitation of DNA and RNA. The broad dynamic range of the chemiluminescent detection assay simplifies the selection of cycling and concentration parameters critical to harnessing the quantit...

journal_title：Methods in enzymology

authors： Martin CS,Voyta JC,Bronstein I

更新日期：2000-01-01 00:00:00

abstract：:The remarkable success of cancer immunotherapies, especially the checkpoint blocking antibodies, in a subset of patients has reinvigorated the study of tumor-immune crosstalk and its role in heterogeneity of response. High-throughput sequencing and imaging technologies can help recapitulate various aspects of the tumo...

journal_title：Methods in enzymology

authors： Bhinder B,Elemento O

更新日期：2020-01-01 00:00:00

abstract：:Measurement of catalysis by MnSOD using direct observation of the UV absorbance of superoxide allows determination of steady-state catalytic constants. Stabilizing superoxide in aprotic solvents such as dimethyl sulfoxide permits the use of stopped-flow spectrophotometry, although significant information is lost in th...

journal_title：Methods in enzymology

authors： Silverman DN,Nick HS

更新日期：2002-01-01 00:00:00

abstract：:Cells belonging to the monocyte/macrophage lineage are in general highly resistant to peroxynitrite. Resistance is not dependent on the scavenging of peroxynitrite itself, or of other secondary reactive species, but is rather associated with the prompt activation of a survival signaling leading to the prevention of to...

journal_title：Methods in enzymology

authors： Cantoni O,Tommasini I,Cerioni L

更新日期：2008-01-01 00:00:00

abstract：:The ability of a systemically administered gene therapy vector to exhibit extended circulation lifetimes, accumulate at a distal tumor site, and enable transgene expression is unique to SPLP. The flexibility and low toxicity of SPLP as a platform technology for systemic gene therapy allows for further optimization of ...

journal_title：Methods in enzymology

authors： Fenske DB,MacLachlan I,Cullis PR

更新日期：2002-01-01 00:00:00

abstract：:Site-directed mutagenesis is a PCR-based method to mutate specified nucleotides of a sequence within a plasmid vector. This technique allows one to study the relative importance of a particular amino acid for protein structure and function. Typical mutations are designed to disrupt or map protein-protein interactions,...

journal_title：Methods in enzymology

authors： Bachman J

更新日期：2013-01-01 00:00:00

abstract：:Classifications and characterizations of specific proteins, such as enzymes, not only allow us to understand biosynthetic and metabolic pathways but they also help to drive our understanding of protein structure and function. How those characterizations are evaluated, however, may change our interpretations and lead u...

journal_title：Methods in enzymology

authors： Bradshaw HB,Leishman E

更新日期：2017-01-01 00:00:00

abstract：:Extracellular signals dictate the biological processes of neural stem cells (NSCs) both in vivo and in vitro. The intracellular response elicited by these signals is dependent on the context in which the signal is received, which in turn is decided by previous and concurrent signals impinging on the cell. A synthesis ...

journal_title：Methods in enzymology

authors： Rajan P,Snyder E

更新日期：2006-01-01 00:00:00

abstract：:The ability to obtain large amounts of genomic sequence for organisms and high throughput technology has led to a change in the thrust of research at the level of chromosomes in animals. In the past chromosomal analysis of animals was focused on gross changes such as inversions, translocations, and deletions for both ...

journal_title：Methods in enzymology

authors： DeSalle R,Gregory TR,Johnston JS

更新日期：2005-01-01 00:00:00

abstract：:This article describes the methods and techniques used to produce mutagenized mice to conduct high-throughput forward genetic screens for circadian rhythm mutants in the mouse. In particular, we outline methods to safely prepare and administer the chemical mutagen N-nitroso-N-ethylurea (ENU) to mice. We also discuss t...

journal_title：Methods in enzymology

authors： Siepka SM,Takahashi JS

更新日期：2005-01-01 00:00:00

abstract：:Embryonic stem (ES) cells are cells derived from the inner cell mass of a blastocyst stage embryo. These self-renewing multipotent cells are able to differentiate to the three embryonic germ layers, the endoderm, ectoderm, and mesoderm, and are thus able to produce virtually all cell types. The ES cell capacity to gen...

journal_title：Methods in enzymology

authors： Goldberg-Cohen I,Beck G,Ziskind A,Itskovitz-Eldor J

更新日期：2006-01-01 00:00:00

abstract：:Linkers play essential roles in the engineering of fusion proteins, and have been extensively demonstrated to affect protein properties such as expression level, solubility, and biological functions. For linker design and optimization, one of the key factors is the flexibility or rigidity of linkers, which describes t...

journal_title：Methods in enzymology

authors： Huang Z,Zhang C,Xing XH

更新日期：2021-01-01 00:00:00

abstract：:The Ras-regulatory network is a loosely defined composition of numerous Ras family members and effector pathways that couple to critical cell-regulatory processes. Investigators are increasingly turning to RNAi-mediated inhibition of gene expression as an effective tool to help generate authentic portraits of Ras prot...

journal_title：Methods in enzymology

authors： Whitehurst AW,White MA

更新日期：2006-01-01 00:00:00

abstract：:Human solid tumors are invariably less well-oxygenated than the normal tissues from which they arose. This so-called tumor hypoxia leads to resistance to radiotherapy and anticancer chemotherapy as well as predisposing for increased tumor metastases. In this chapter, we examine the resistance of tumors to radiotherapy...

journal_title：Methods in enzymology

authors： Brown JM

更新日期：2007-01-01 00:00:00