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Cell Pseudopodia and Purification Assays

Pseudopodia assays are ideal for screening pharmacological agents that stimulate or inhibit pseudopodia formation, and therefore, impact cell chemotaxis. The Quantitative Pseudopodia Assay is a novel system designed to specifically isolate and quantitate extending or retracting pseudopodia from the cell body. The assay is a quick and efficient system to assess the effects of various natural or synthetic compounds that specifically impact the microenvironment of the pseudopodial organelle including signaling processes, integrins or other adhesion receptors, and unique gene products. It provides a novel method to purify the pseudopodia proteins for further biochemical characterization.

Gradient Sensing and Isolation of Leading Pseudopodia

	A. Reducing SDS-PAGE of 2.5 µg purified chicken tenascin (Millipore cat. no. CC115) illustrating a major band at 220 kDa, with doublet at 200 kDa.
	B. Tenascin is known to play a role in development of the CNS and mesenchymal-derived organs, exist in vasculature of tumors in adults and function in cell adhesion. SDS-PAGE of purified Human Tenascin (Millipore cat. no. CC065)
	C. Reducing SDS-PAGE of 2.5 µg purified restrictin protein (Millipore cat. no. CC116) illustrating a major band at 180 kDa.

Applications

• Measure directional pseudopodia extension
• Analyze spatial and temporal changes in focal adhesion, microtubules and the actin/myosin cytoskeleton
• Monitor gradient sensing behavior
• Perform quantitative time-lapse imaging of pseudopodia extension and retraction
• Harvest pseudopodia proteins and isolate trailing cell body proteins
• Run functional assays on purified pseudopodia proteins

Quantitative Pseudopodia Assay Kit Components

• Pseudopodia Quantification Inserts: 1 X 24 well plate containing 12 transwells
• Pseudopodia Stain Solution
• Pseudopodia Stain Extraction Buffer
• Forceps
• Cotton Swabs

Pseudopodia Purification Kit Components

• Pseudopodia Purification Inserts:1 X 6 well plate containing 6 transwells
• Western Blot Lysis Buffer
• Immunoprecipitation Buffer
• Cell Scrapers
• Forceps
• Cotton Swabs

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