MilliporeSigma™ LC3-II Enrichment Kit (Western Blot)

Description

Includes

LC3-II Enrichment Component Box

• Autophagy Reagent A: One vial, lyophilized
• 10x Enrichment Buffer: One bottle containing 10mL 10x Enrichment Buffer
• 5x Assay Buffer: One bottle containing 15mL 5x Assay Buffer
• 10x RIPA Lysis Buffer: One bottle containing 1mL 10x RIPA Lysis Buffer

Western Blot Detection Component Box

• Protease Inhibitor Cocktail: One vial containing 20 μL of 1000x Protease Inhibitor Cocktail
• Anti-LC3 antibody: One vial containing 20μL of 1000x anti-LC3 rabbit monoclonal antibody
• Anti-TOMM22 antibody: One vial containing 20μL of 1000x anti-TOMM22 mouse monoclonal antibody
• Goat anti-Mouse IgG HRP, Secondary Antibody: One vial containing 30μL of 2000X goat anti-mouse HRP conjugate
• Goat anti-Rabbit IgG HRP, Secondary Antibody: One vial containing 30μL of 2000X goat anti-rabbit HRP conjugate

Requires

• Pipettors and tips capable of accurately measuring 1-1000μL
• Graduated serological pipettes
• Microcentrifuge tubes
• Mechanical vortex
• Rocker
• Distilled or deionized water
• 1x Dulbecco's Phosphate Buffered Saline (D-PBS)
• 1x Tris-buffered saline containing 0.05% Tween-20 (TBST)
• 3% Non-fat dry milk diluted in TBST (TBST-MILK)
• Earle's Balanced Salt Solution (EBSS)

Enables sensitive and accurate quantification of autophagosome density by utilizing selective permeabilization procedure that removes cytosolic LC3-I and retains autophagosome-bound LC3-II

• Enables sensitive and accurate quantification of autophagosome density by utilizing a selective permeabilization procedure that removes cytosolic LC3-I and retains autophagosome-bound LC3-II
• Reveals quantities of LC3-II, without interference from LC3-I, by Western blotting analysis
• The kit also includes a lysosomal inhibitor, to prevent degradation of LC3-II that occurs upon fusion of the autophagosome with the lysosome
• An antibody against a mitochondrial outer membrane protein TOMM22 is included for use as a control for specificity of permeabilization, to ensure that organelles remain intact after treatment

Specifications

• Content And Storage: The kit may be used for up to 4 months after receipt. Avoid repeated freeze/thaw cycles, and aliquot if necessary
• For Use With (Application): Western Blotting
• Quantity: 15 Assays