Comparative Proteomics Kit II: Western Blot Module

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Overview

Students use western blotting to specifically identify myosin light chain from the hundreds of other proteins that comprise the muscle cell extracts of closely and distantly related species of fish.
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Description

Description

The Comparative Proteomics Kit II: Western Blot Module is the second part of a two-part laboratory. In the first part of this laboratory (Comparative Proteomics Kit I: Protein Profiler Module), students use sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to generate protein profiles and to visualize the unique array of proteins comprising muscle tissues from different fish. In this second part of the laboratory, students apply western blotting techniques to their polyacrylamide gel results to specifically identify myosin light chain from the hundreds of proteins that comprise the muscle cell extracts of closely and distantly related species of fish.

Features and Benefits

  • Applies immunology
  • Teaches students to use antibodies as detection tools
  • Serves as a laboratory extension for the Comparative Proteomics I Kit: Protein Profiler Module

How It Works

The protocol involves transferring the protein bands from polyacrylamide gels to a membrane and then using an anti-myosin light chain antibody to precisely identify which protein in each species' profile is myosin light chain. Using Internet-based bioinformatics databases, students can compare their experimentally determined results to actual protein sequence data derived from DNA and RNA sequences and consider whether variations in myosin among species are due to "genetic" or "epigenetic" factors.

More Information

  • Four-session laboratory activity, 45 min per session
  • Provides sufficient materials for eight student workstations, up to four students per workstation

Specifications

Specifications

Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)
 
Primary antibody (anti-myosin light chain mouse monoclonal), lyophilized
1
Secondary antibody (goat anti-mouse polyclonal antibody conjugated to horseradish peroxidase (HRP), lyophilized
1
HRP color detection reagent A
1
HRP color detection reagent B
1
10x Tris-glycine
1 L
Non-fat dry milk blocker
1
10x phosphate buffered saline (PBS)
2 x 100 ml
10% Tween 20
5 ml
Nitrocellulose sheets, 0.45 µm
8
Blotting paper sheets
16
Reagent tubes
25
Curriculum, including teacher's guide, student manual, and graphic quick guide
1

Ordering

Ordering

items

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1662800EDU
Western blot classroom study kit, includes antibodies, color detection reagents, 10x TG buffer, 10x PBS, 10% Tween, nitrocellulose, curriculum, and more, for 32 students; education use only
166-2850EDU_tn.jpg
1662850EDU
Comprehensive protein classroom study kit, includes Protein Profiler Module (1662700EDU) and Western Blot Module (1662800EDU), for 32 students; education use only
166-2875EDU_tn.jpg
1662875EDU
Comprehensive protein classroom study kit, includes Protein Profiler Module (1662700EDU), Western Blot Module (1662800EDU), Trans-Blot® Turbo Mini Nitrocellulose Transfer Packs (10), 4–20% Mini-PROTEAN® TGX Stain-Free Gels (10), application note, for 32 students; education use only

Refills

Refills

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1662801EDU
Consumables refill reagent kit for use with Comparative Proteomics Kit II: Western Blot Module (1662800EDU), includes antibodies and color detection reagents; education use only
166-2804EDU_tn.jpg
1662804EDU
200 μg, lyophilized, primary antibody (anti-myosin light chain antibody); education use only
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(Discontinued) Goat X Mouse-HRP
1721011EDU
2 ml, EIA-grade, enzyme-antibody conjugate; education use only
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1706431EDU
Colorimetric detection for western blotting kit, includes premixed 4CN, hydrogen peroxide solutions, color development buffer; makes 1 L color development solution; education use only
166-2807EDU_tn.jpg
1662807EDU
0.45 µm, 7 x 8.4 cm, nitrocellulose membrane (8 sheets)/filter paper (16 sheets) sandwich; education use only
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1662876EDU
Application note for western blotting, describes rapid blotting using the Trans-Blot® Turbo Transfer System and V3 Western Workflow (stain-free blotting) methods; education use only Use Add to Cart to order a printed copy or download a free PDF.

Documents

Documents

Number Description Options
M1662876EDU
Bio-Rad Explorer™ Rapid Blotting + V3 Western Workflow (Stain-Free Rapid Blotting) Application Note, Rev A
10004531
Instruction Manual, Comparative Proteomics Kit II: Western Blot Module, Bio-Rad Explorer Rev E
5478
Instruction Manual, Comparative Proteomics Kit II: Western Blot Module Quick Guide, Bio-Rad Explorer™, Rev A
6933
Fast Electrophoresis and Western Blotting Flier