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University of East Anglia

4 ARTICLES PUBLISHED IN JoVE

Biochemistry

Species Determination and Quantitation in Mixtures Using MRM Mass Spectrometry of Peptides Applied to Meat Authentication

Yvonne Gunning¹, Andrew D. Watson¹, Neil M. Rigby², Mark Philo¹, Joshua K. Peazer^1,3, E. Kate Kemsley¹

¹Analytical Sciences Unit, Institute of Food Research, ²Institute of Food Research, ³School of Chemistry, University of East Anglia

We present a protocol for identifying and quantifying the components in mixtures of species possessing similar proteins. Mass spectrometry detects peptides for identification, and gives relative quantitation by ratios of peak areas. As a tool food for fraud detection, the method can detect 1% horse in beef.

JoVE Core

Spin Saturation Transfer Difference NMR (SSTD NMR): A New Tool to Obtain Kinetic Parameters of Chemical Exchange Processes

María Teresa Quirós¹, Colin Macdonald^1,2, Jesús Angulo², María Paz Muñoz¹

¹School of Chemistry, University of East Anglia, ²School of Pharmacy, University of East Anglia

A detailed protocol describing the SSTD NMR method is presented here to help new users apply this new method to obtain the kinetic parameters of their own systems undergoing chemical exchange.

Behavior

The (Spatial) Memory Game: Testing the Relationship Between Spatial Language, Object Knowledge, and Spatial Cognition

Harmen B. Gudde¹, Debra Griffiths¹, Kenny R. Coventry¹

¹School of Psychology, University of East Anglia

We present a protocol to explore the relationship between spatial language production, spatial memory, and object knowledge. The procedure allows experimental manipulation of, and control over, conditions of object knowledge, language at instruction, and physical location, thus teasing apart cognitive and linguistic models describing interactions between these variables.

Developmental Biology

Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids

Deborah A. Goldspink¹, Zoe J. Matthews², Elizabeth K. Lund¹, Tom Wileman², Mette M. Mogensen¹

¹School of Biological Sciences, University of East Anglia, ²Norwich Medical School, University of East Anglia

We present protocols for isolation of intestinal 3D structures from in vivo tissue and in vitro basement matrix embedded organoids, and detail different fixation and staining protocols optimized for immuno-labeling of microtubule, centrosomal, and junctional proteins as well as cell markers including the stem cell protein Lgr5.