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Volume:4  Issue:19

October 05, 2014

Cancer Biology

Hypoxia Studies with Pimonidazole in vivo

Authors: Kristina Y. Aguilera
Kristina Y. AguileraAffiliation: Department of Surgery, Hamon Center for Therapeutic Oncology Research, Dallas, USA
Bio-protocol author page: a1691
 and Rolf A. Brekken
Rolf A. BrekkenAffiliation 1: Department of Surgery, Hamon Center for Therapeutic Oncology Research, Dallas, USA
Affiliation 2: Department of Pharmacology, UT Southwestern Medical Center, Dallas, USA
For correspondence: rolf.brekken@utsouthwestern.edu
Bio-protocol author page: a1692
date: 10/5/2014, 8214 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1254 

[Abstract] Therapy-induced hypoxia drives changes in the tumor microenvironment that contribute to the poor response to therapy. Hypoxia is capable of driving the expression and/or activation of specific signaling cascades (e.g., c-Met, Axl, CTGF), the recruitment ...

Article identifier: bio-protocol.org/e1254

Immunology

Coagulation Assay

Authors: Praveen Papareddy
Praveen PapareddyAffiliation: Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University Biomedical Center, Lund, Sweden
For correspondence: praveen.papareddy@med.lu.se
Bio-protocol author page: a1672
Martina Kalle
Martina KalleAffiliation: Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University Biomedical Center, Lund, Sweden
Bio-protocol author page: a1673
 and Artur Schmidtchen
Artur SchmidtchenAffiliation: Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University Biomedical Center, Lund, Sweden
Bio-protocol author page: a1674
date: 10/5/2014, 3544 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1247 

[Abstract] Clotting times can be measured by using citrate plasma. The intrinsic pathway of coagulation is measured by the activated partial thromboplastin time (aPTT), the extrinsic pathway of coagulation, monitored by measuring the prothrombin time (PT), and ...

Article identifier: bio-protocol.org/e1247

Isolation and in vivo Transfer of Antigen Presenting Cells

Authors: Pooja Arora
Pooja AroraAffiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, NY, USA
Bio-protocol author page: a1709
Shalu Sharma Kharkwal
Shalu Sharma KharkwalAffiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, NY, USA
Bio-protocol author page: a1710
 and Steven A. Porcelli
Steven A. PorcelliAffiliation: Department of Medicine, Albert Einstein College of Medicine, NY, USA
For correspondence: steven.porcelli@einstein.yu.edu
Bio-protocol author page: a1711
date: 10/5/2014, 3795 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1260 

[Abstract] Transfer of antigen presenting cells in vivo is a method used by immunologists to examine the potency of antigen presentation by a selected population of cells. This method is most commonly used to analyze presentation of protein antigens to MHC class ...

Article identifier: bio-protocol.org/e1260

Microbiology

Chromogenic Substrate Assay for Determining the Activity of Plasma Kallikrein

Authors: Praveen Papareddy
Praveen PapareddyAffiliation: Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University Biomedical Center, Lund, Sweden
For correspondence: praveen.papareddy@med.lu.se
Bio-protocol author page: a1672
Martina Kalle
Martina KalleAffiliation: Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University Biomedical Center, Lund, Sweden
Bio-protocol author page: a1673
 and Artur Schmidtchen
Artur SchmidtchenAffiliation: Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University Biomedical Center, Lund, Sweden
Bio-protocol author page: a1674
date: 10/5/2014, 2669 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1246 

[Abstract] The activation of the intrinsic pathway takes place at negatively charged surfaces, such as bacteria, and involves activation of cogulation Factor XII, which then leads to the activation of plasma kallikrein (PK) and coagulation Factor XI. To determine ...

Article identifier: bio-protocol.org/e1246

Construction of Glycine Oxidase Mutant Libraries by Random Mutagenesis, Site Directed Mutagenesis and DNA Shuffling

Author: Tao Zhan
Tao ZhanAffiliation 1: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
Affiliation 2: Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin, China
For correspondence: zhan_t@tib.cas.cn
Bio-protocol author page: a1687
date: 10/5/2014, 2775 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1252 

[Abstract] Glyphosate, a broad spectrum herbicide widely used in agriculture all over the world, inhibits 5-enolpyruvylshikimate-3-phosphate synthase in the shikimate pathway, and glycine oxidase (GO) has been reported to be able to catalyze the oxidative deamination ...

Article identifier: bio-protocol.org/e1252

Expression and Purification of the Thermus thermophilus Argonaute Protein

Authors: Daan C. Swarts
Daan C. SwartsAffiliation: Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, The Netherlands
For correspondence: daan.swarts@wur.nl
Bio-protocol author page: a1688
Matthijs M. Jore
Matthijs M. JoreAffiliation: Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, The Netherlands
Bio-protocol author page: a1689
 and John van der Oost
John van der OostAffiliation: Department of Agrotechnology and Food Sciences, Wageningen University, Wageningen, The Netherlands
Bio-protocol author page: a1690
date: 10/5/2014, 2879 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1253 

[Abstract] The Argonaute protein of Thermus thermophilus (TtAgo) has recently been studied in detail. For its in vitro characterization, TtAgo was purified after heterologous expression in Escherichia coli (E. coli). As TtAgo expression is toxic, a tightly controlled ...

Article identifier: bio-protocol.org/e1253

Neuroscience

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Novel Object Recognition for Studying Memory in Mice

Authors: Tzyy-Nan Huang
Tzyy-Nan HuangAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: eugene02@gate.sinica.edu.tw
Bio-protocol author page: a1680
 and Yi-Ping Hsueh
Yi-Ping HsuehAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: yph@gate.sinica.edu.tw
Bio-protocol author page: a1681
date: 10/5/2014, 11837 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1249 

[Abstract] Memory tests are important indexes of the brain functions for rodents behavior assay. Many memory tasks require external forces (e.g. electric shocks) or intrinsic forces (e.g. hunger and thirsty) to trigger the responses. Under those conditions, rodents ...

Article identifier: bio-protocol.org/e1249

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Two-choice Digging Task in Mouse for Studying the Cognitive Flexibility

Authors: Hsiu-Chun Chuang
Hsiu-Chun ChuangAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: hcchuang@imb.sinica.edu.tw
Bio-protocol author page: a1682
Tzyy-Nan Huang
Tzyy-Nan HuangAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: eugene02@gate.sinica.edu.tw
Bio-protocol author page: a1680
 and Yi-Ping Hsueh
Yi-Ping HsuehAffiliation: Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan
For correspondence: yph@gate.sinica.edu.tw
Bio-protocol author page: a1681
date: 10/5/2014, 3534 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1250 

[Abstract] Cognitive flexibility, the higher-order cognition involving reversal learning, has been defined as having the ability to shift one’s previous thoughts or actions to new situations depending on situational demands. Studies of neuropsychiatric disorders ...

Article identifier: bio-protocol.org/e1250

In utero Electroporation of the Embryonic Mouse Retina

Authors: Francisco Nieto-Lopez
Francisco Nieto-LopezAffiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Madrid, Spain
Bio-protocol author page: a1694
 and Luisa Sanchez-Arrones
Luisa Sanchez-ArronesAffiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Madrid, Spain
For correspondence: luisasanchez@cbm.uam.es
Bio-protocol author page: a1693
date: 10/5/2014, 4657 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1255 

[Abstract] This protocol is useful to manipulate gene expression in the embryonic retina and compare the result with the contralateral non electroporated retina. In addition, the electroporation of a membrane or cytoplasmic tagged GFP allows to determine the effects ...

Article identifier: bio-protocol.org/e1255

Plant Science

Analysis of Total Se Content in Rice

Authors: Lianhe Zhang
Lianhe ZhangAffiliation: Agricultural Faculty, Henan University of Science and Technology, Luoyang, China
For correspondence: lhzhang2007@126.com
Bio-protocol author page: a1675
Feiyan Yu
Feiyan YuAffiliation: Agricultural Faculty, Henan University of Science and Technology, Luoyang, China
Bio-protocol author page: a1676
Kun Deng
Kun DengAffiliation: Agricultural Faculty, Henan University of Science and Technology, Luoyang, China
Bio-protocol author page: a1677
Bin Hu
Bin HuAffiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a1678
 and Chengcai Chu
Chengcai ChuAffiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China
For correspondence: ccchu@genetics.ac.cn
Bio-protocol author page: a1679
date: 10/5/2014, 2320 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1248 

[Abstract] Total Se content in rice is normally low and it is difficult to determine it exactly because of Se volatilization and pollution during the digestion process. In this method, rice sample is digested thoroughly and Se volatilization is reduced greatly ...

Article identifier: bio-protocol.org/e1248

Pea Aphid Survival Assays on Arabidopsis thaliana

Authors: David C. Prince
David C. PrinceAffiliation: Department of Cell and Development Biology, John Innes Center, Norwich Research Park, Norwich, UK
Bio-protocol author page: a1683
Sam T. Mugford
Sam T. MugfordAffiliation: Department of Cell and Development Biology, John Innes Center, Norwich Research Park, Norwich, UK
Bio-protocol author page: a1684
Thomas R. Vincent
Thomas R. VincentAffiliation: Department of Metabolic Biology, John Innes Center, Norwich Research Park, Norwich, UK
Bio-protocol author page: a1685
 and Saskia A. Hogenhout
Saskia A. HogenhoutAffiliation: Department of Cell and Development Biology, John Innes Center, Norwich Research Park, Norwich, UK
For correspondence: saskia.hogenhout@jic.ac.uk
Bio-protocol author page: a1686
date: 10/5/2014, 2819 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1251 

[Abstract] Aphids are phloem-feeding insects that successfully colonize specific host plant species. Aphid performance on a given plant is commonly measured by assessing fecundity of an aphid species that is adapted to the host. However, this approach may not reveal ...

Article identifier: bio-protocol.org/e1251

Carotenoid Extraction and Quantification from Capsicum annuum

Authors: Richard D. Richins
Richard D. RichinsAffiliation: Plant and Environmental Sciences, New Mexico State University, Las Cruces, USA
Bio-protocol author page: a1695
James Kilcrease
James KilcreaseAffiliation: Plant and Environmental Sciences, New Mexico State University, Las Cruces, USA
Bio-protocol author page: a1696
Laura Rodgriguez-Uribe
Laura Rodgriguez-UribeAffiliation: Plant and Environmental Sciences, New Mexico State University, Las Cruces, USA
Bio-protocol author page: a1697
 and Mary A. O’Connell
Mary A. O’ConnellAffiliation: Plant and Environmental Sciences, New Mexico State University, Las Cruces, USA
For correspondence: moconnel@nmsu.edu
Bio-protocol author page: a1698
date: 10/5/2014, 4001 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1256 

[Abstract] Carotenoids are ubiquitous pigments that play key roles in photosynthesis and also accumulate to high levels in fruit and flowers. Specific carotenoids play essential roles in human health as these compounds are precursors for Vitamin A; other specific ...

Article identifier: bio-protocol.org/e1256

Endogenous ABA Extraction and Measurement from Arabidopsis Leaves

Authors: Ning Liu
Ning LiuAffiliation: School of Biological Sciences, University of Nebraska, Lincoln, USA
For correspondence: nliu2@unl.edu
Bio-protocol author page: a1699
Yong Ding
Yong DingAffiliation 1: School of Life Sciences, University of Science & Technology of China, Hefei, China
Affiliation 2: Center for Biotechnology and Center for Plant Science Innovation, University of Nebraska, Lincoln, USA
Bio-protocol author page: a1700
Michael Fromm
Michael FrommAffiliation: School of Life Sciences, University of Science & Technology of China, Hefei, China
Bio-protocol author page: a1701
 and Zoya Avramova
Zoya AvramovaAffiliation: School of Biological Sciences, University of Nebraska, Lincoln, USA
Bio-protocol author page: a1702
date: 10/5/2014, 3669 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1257 

[Abstract] The endogenous messenger, the phytohormone abscisic acid (ABA) plays a major role in plant’s adaption to drought, salinity, cold and other abiotic stresses. In addition to abiotic stress signaling, ABA is involved also in developmental regulation and ...

Article identifier: bio-protocol.org/e1257

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Agrobacterium tumefaciens-mediated Transformation of Walnut (Juglans regia)

Authors: Sriema L. Walawage
Sriema L. WalawageAffiliation: Department of Plant Sciences, University of California, Davis, USA
Bio-protocol author page: a1703
Charles A. Leslie
Charles A. LeslieAffiliation: Department of Plant Sciences, University of California, Davis, USA
Bio-protocol author page: a1704
Matthew A. Escobar
Matthew A. EscobarAffiliation: Department of Biological Sciences, California State University San Marcos, San Marcos, USA
Bio-protocol author page: a1587
 and Abhaya M. Dandekar
Abhaya M. DandekarAffiliation: Department of Plant Sciences, University of California, Davis, USA
For correspondence: amdandekar@ucdavis.edu
Bio-protocol author page: a1705
date: 10/5/2014, 3539 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1258 

[Abstract] Like many woody plant species, walnut (Juglans regia) can be difficult to genetically transform and regenerate. However, somatic embryos have been used successfully for over two decades as a target tissue for transformation and regeneration of transgenic ...

Article identifier: bio-protocol.org/e1258

Localisation and Quantification of Reactive Oxygen Species and Nitric Oxide in Arabidopsis Roots in Response to Fungal Infection

Authors: Kapuganti J. Gupta
Kapuganti J. GuptaAffiliation: Department of Plant Sciences, University of Oxford, Oxford, UK
For correspondence: jagadis.kapuganti@plants.ox.ac.uk
Bio-protocol author page: a1706
Yariv Brotman
Yariv BrotmanAffiliation: Max-Planck-Institute of Molecular Plant Physiology, Golm-Potsdam, Germany
Bio-protocol author page: a1707
 and Luis A. J. Mur
Luis A. J. MurAffiliation: Institute of Environmental and Rural Science, Aberystwyth University, Aberystwyt, UK
Bio-protocol author page: a1708
date: 10/5/2014, 3734 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1259 

[Abstract] Nitric oxide and reactive oxygen species have emerged as important signalling molecules in plants. The half-lives of NO and ROS are very short therefore rapid and precise measurements are required for the understanding biological roles of these redox ...

Article identifier: bio-protocol.org/e1259