Protein Expression and Purification Series

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Overview

Protein Expression and Purification Checklist
Protein Expression and Purification Series Inventory Checklist

Lab contains sufficient materials for 12 student workstations for the Centrifugation Purification Process and four student workstations for the Hand-packed and Prepacked Purification Processes

Centrifugation Purification Process (catalog #166-5040EDU)
Kit Components Quantity  
Growth and Expression Module 1  
SDS-PAGE Electrophoresis Module 1  
Centrifugation Purification Module 1  
DHFR Enzymatic Assay Module 1  
Hand-packed Purification Process (catalog #166-5045EDU)
Kit Components Quantity  
Growth and Expression Module 1  
SDS-PAGE Electrophoresis Module 1  
Hand-packed Purification Module 1  
DHFR Enzymatic Assay Module 1  
Prepacked Purification Process (catalog #166-5050EDU)
Kit Components Quantity  
Growth and Expression Module 1  
SDS-PAGE Electrophoresis Module 1  
Prepacked Purification Module 1  
DHFR Enzymatic Assay Module 1  
Materials Required But Not Supplied
Chemicals Centrifugation Purification BioLogic™ LP/BioLogic DuoFlow™ Purification
Dry ice 250 g 250 g
Ethanol 500 ml 500 ml
Plastics and Consumables Centrifugation Purification BioLogic LP/BioLogic DuoFlow Purification
100–1,000 μl pipet tips, standard style (catalog #223-9350EDU) 12 boxes 4 boxes
2–200 μl pipet tips, standard style (catalog #223-9347EDU) 12 boxes 4 boxes
Parafilm sealing film 1 1
trUView™ cuvettes (catalog #170-2510EDU) 1 box of 50 1 box of 50
or or or
quartz submicrovolume cuvettes (catalog #170-2505EDU) 1–4 cuvettes 1–4 cuvettes
1.5 ml standard disposable polystyrene cuvettes (catalog #223-9955EDU) 1 box of 100 1 box of 100
or or or
quartz cuvettes (catalog #170-2502EDU) N/A 1–4 cuvettes
10 ml syringes 12 16
22 gauge syringe needles 12 4
Fraction collection tubes (catalog #223-9751EDU) N/A 200 tubes
Glassware Centrifugation Purification BioLogic LP/BioLogic DuoFlow Purification
500 ml Erlenmeyer flasks N/A 4
1 L Erlenmeyer flasks or autoclavable bottles 2 1
Beakers for dry ice/ethanol bath 1–12 1–4
Temperature Control Equipment Centrifugation Purification BioLogic LP/BioLogic DuoFlow Purification
Microwave oven 1 1
–20°C freezer 1 1
Incubation oven (catalog #166-0501EDU) 1–2* 1
Tube roller (catalog #166-0711EDU) 1–2* N/A
Mini rocker (catalog #166-0710EDU) 1** 1
Shaking incubator or shaking water bath 1 N/A
capable of holding 4 x 500 ml flasks N/A 1
Dry bath (catalog #166-0562EDU) or water bath 1 1
* Initial overnight cell cultures and cell subcultures/induced cells for Centrifugation Purification are prepared in 50 ml sterile conical tubes. Incubation at 37°C with some form of mixing is required. This incubation can be accomplished in either a temperature controlled shaking incubator, a shaking water bath, or using tube rollers in an incubation oven set at 37°C. The Bio-Rad tube roller can hold six 50 ml sterile conical tubes; therefore, if 12 workstations are being run, two tube rollers, each in its own incubation oven, would be required.

** For the Centrifugation Purification protocol, during the binding of the cell lysate to the Profinity™ IMAC Ni-Resin, a tube roller can be used for end-over-end mixing of the resin and lysate. If a tube roller is not available, the mixing can be accomplished by using a mini rocker. The mini rocker can be further used for rinsing, staining, and destaining of SDS-PAGE gels.
Other Equipment Centrifugation Purification BioLogic LP/BioLogic DuoFlow Purification
Spectrophotometer capable of reading to three decimal places (catalog #170-2525EDU) 1 1
100–1,000 μl adjustable micropipet (catalog #166-0508EDU, 166-0553EDU) 12 4
20–200 μl adjustable micropipet (catalog #166-0507EDU, 166-0552EDU) 12 4
2–20 μl adjustable micropipet (catalog #166-0506EDU, 166-0551EDU) 12 4
Pipet pump or filler 12 4
10 ml serological pipets 4 4
Power supply (catalog #164-5050EDU) 3–12 3–12
Vertical electrophoresis chambers (catalog #165-8004EDU, 165-8005EDU) 3–12 1–4
Gel documentation system (catalog #170-8170EDU, 170-8270EDU) 1 1
Microcentrifuge with variable speed setting capable of 16,000 x g (catalog #166-0602EDU) 1 1
Centrifuge capable of 16,000 x g with rotors that hold 250 ml centrifuge bottles and 30–50 ml centrifuge tubes N/A 1
Centrifuge bottles (250 ml) capable of withstanding 4,500 x g N/A 4
Centrifuge tubes (30–50 ml) capable of withstanding 16,000 x g N/A 4
Hand-packed Column Process Only Quantity  
Flow Adaptor, 1.0 cm column ID, 1–7 cm functional length (catalog #738-0014EDU) 1 per instrument  
Equipment Necessary if Using a BioLogic DuoFlow System
(catalog #760-0047EDU or 760-0037EDU)		 Quantity  
Fittings to convert luer to ¼–28 (catalog #732-0113EDU) 1 per instrument  
5 ml injection loop (catalog #750-0497EDU) 1 per instrument  
Miscellaneous Centrifugation Purification BioLogic LP/BioLogic DuoFlow Purification
Marking pens 12 4
Storage boxes for microcentrifuge tubes (catalog #166-0482EDU) 3 1
Green racks for microcentrifuge tubes (catalog #166-0481EDU) 12 4
Optional Materials Centrifugation Purification BioLogic LP/BioLogic DuoFlow Purification
Autoclave 1 1
Vortexer (catalog #166-0610EDU) 1 N/A
Imaging System (catalog #170-8270EDU with 170-8272EDU) 1 1
Workflow
Workflow
Protein Expression Workflow
Webinar
Webinar
Jim DeKloe

From industrial enzymes to cancer therapy — protein purification is essential!
May 11, 2011
Jim DeKloe
Solano Community College, Fairfield, California

Register to play video

This webinar shows you how easy it is to teach the core process of protein expression and purification using the clear and concise modules in the Protein Expression and Purification Series (available summer 2011). Give your students hands-on experience and the confidence they need to work with proteins. The scalability of this affinity purification process ensures that techniques and content can be adapted to match the goals of protein educators from basic lessons to an advanced college level course.

Discover more about this series, including how dihydrofolate reductase, DHFR, is a target for certain cancer treatments and how the protein expression and purification process can be relevant to the world of biomanufacturing.

Speaker's bio:
Jim DeKloe has been a professor at Solano Community College in the San Francisco Bay area since 1990, and he currently serves as the director of the Industrial Biotechnology Program. This program trains students in the skills required to work in biotechnology companies, particularly those located in the biotech manufacturing cluster in Solano County between San Francisco and Sacramento. Jim also serves as the regional director of the Southwest Region of Bio-Link, the nationwide consortium of community and technical colleges that teach biotech. During a yearlong sabbatical, Jim worked in the manufacturing department of biotech pioneer Genentech, Inc., to gain real-world knowledge of biomanufacturing processes and needs. As a consultant, he helped the biotech giant Amgen redesign its training programs for biomanufacturing technicians and quality assurance associates. In 2000, the Association of Community College Trustees chose Jim as the Distinguished Faculty Member of the Pacific Region, which includes the western continental United States, Hawaii, Alaska, Canada, and Guam.
PowerPoint Presentation for educational use
PowerPoint Presentation for Educational Use

Protein Expression and Purification Series

This PowerPoint presentation is intended for educational use only. The presentation provides educators with background on the Protein Expression and Purification Series and includes some instructions on the series activities.

DNA Fingerprinting Kit Download PowerPoint presentation		 Protein Expression and Purification Series

The Protein Expression and Purification Series provides students with the opportunity to express, affinity purify, and evaluate enzymatic activity of DHFR. DHFR is an essential protein in nucleic acid synthesis and was one of the first targets for cancer chemotherapy. The scalability of this particular affinity purification process provides an adaptable set of techniques and content to match the goals of the beginning protein educator up to an advanced college level course in biomanufacturing. The series provides a fully developed and ready-to-go lab course, including relevant background, protocols that work, and student assessment.

Features and Benefits

Designed by undergraduate and community college educators, this course was specifically developed to ensure success for you and your students. The modular nature of the series allows you to choose from three different purification options depending on your teaching goals and equipment resources:

  • Centrifugation Purification Process – allowing protein purification using a 16K Microcentrifuge
  • Hand-packed Purification Process – for use with chromatographic purification instrumentation and allowing students to pour their own columns
  • Prepacked Purification Process – providing the best experience with chromatographic purification instrumentation through the quality of prepacked cartridges

An Assessment Module is also available to complement the hands-on laboratory activities that the series provides. This unique assessment guide provides ideas for using formative assessment in your class to guide and increase learning while students perform the lab activities. At the end of the lab series use the summative assessment to evaluate the final learning levels of your students. The assessment tool is arranged according to learning levels so that you can choose what questions to modify instruction and evaluate the comprehension of your students.

The Protein Expression and Purification Series allows students to apply molecular biology skills and techniques to complete a real-world research-level project, potentially encouraging them to pursue a career in research.

More Information

  • 3–5 weeks of laboratory activities made up of four modular kits
  • Provides sufficient materials for 12 student workstations for the Centrifugation Purification Process and four student workstations for the Hand-packed and Prepacked Purification Processes, up to four students per workstation

Patent Pending

Protein Expression and Purification Series – Centrifugation Purification Process

166-5040EDU
Biotechnology study kit, includes 4 modules with lab activities from protein expression and purification using spin columns to SDS-PAGE and enzymatic activity analysis, curriculum

List Price:   $708.75
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Protein Expression and Purification Series – Hand-packed Purification Process

166-5045EDU
Biotechnology study kit, includes 4 modules with lab activities from protein expression and purification using hand-packed columns to SDS-PAGE and enzymatic activity analysis, curriculum

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Protein Expression and Purification Series – Prepacked Purification Process

166-5050EDU
Biotechnology study kit, includes 4 modules with lab activities from protein expression and purification using prepacked cartridges to SDS-PAGE and enzymatic activity analysis, curriculum

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Protein Expression and Purification Series Formative and Summative Assessment Module

166-5070EDU
Assessment guide provides ideas for using formative assessment and summative assessment to aid in evaluating the learning levels of your students

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Number Description Options
6160 Biotechnology Explorer Protein Expression and Purification Series Planning Guide, Rev A Click to download
1665067 Instruction Manual, Protein Expression and Purification Series, Biotechnology Explorer, Rev A Please login to download this document