Minute™ Plasma Membrane Protein Isolation Kit
Minute™ plasma membrane (PM) protein isolation kit is a novel true native (detergent and EDTA free) PM protein isolation kit coupled with unique mechanisms of action using our patented technologies. Cells/Tissues are sensitized before passing through a filter that allows cells to travel in a convoluted path. The cell membranes are ruptured during the process without using traditional homogenization or sonication. PM is separated from a mixture of residual un-ruptured cells, nuclei, cytosolic proteins and organelles by subsequent differential and density centrifugations using a desk top micro-centrifuge. Ultracentrifugation is not required. By eliminating unwanted variations caused by manual homogenization or sonication, MinuteTM plasma membrane protein isolation kit enables researchers to obtain consistent results in less than 45 minutes.
Price:$298 (50 preps)
Figure 1. A. SDS-PAGE profiles of total cell lysate (TC) and isolated plasma membrane proteins (PM) from human and rat cultured cells. Lane 1, Human lung cancer cell A549 total cell lysate; Lane 2, Isolated PM of A549; Lane 3, PM of rat REL-6TN cells; Lane 4, Total cell lysate of rat REL-6TN cells.
B. Western blottings : Proteins in A were transferred to nitrocellulose membrane and probed with anti-Na/K ATPase alpha1, a plasma membrane marker (Upstate, Clone 464.6)) and anti-lamin B1, a nuclear envelope marker (ab16048, abcam Cambridge, MA). The specific protein bands were visualized by a color metric substrate Opti-4CN (Bio-RAD).
C. Densitometry measurement of Na/K ATPase alpha1 signals in B (TC vs. PM). Total cell lysates were extracted by Minute Denaturing Total Protein Extraction Kit (SD-001, Invent Biotechnologies, Eden Prairie, MN).
Figure 2 A. SDS-PAGE profiles of isolated total membrane proteins from mouse tissues (T=Total Cell lysate, C=Cytosol Fraction, M= Total Membrane Fraction)
B. Proteins shown in A were transferred to a nitrocellulose membrane and probed with rabbit-anti mouse pan-cadherin (ab6529,abcam, Cambridge, MA), and anti-actin by Western blotting. The specific protein bands were visualized by a color metric substrate Opti-4CN (Bio-RAD). Signals of pan-cadherin (a 130 kda plasma membrane marker) were significantly enhanced in total membrane protein fractions. Total cell lysates were extracted by MinuteTM Denaturing Total Protein Extraction Kit (SD-001, Invent Biotechnologies, Eden Prairie, MN).
Figure 3. A. SDS-PAGE profiles of total membrane protein fraction (TM) and isolated plasma membrane protein fraction from mouse tissues.
B. Western blotting of proteins in A were transferred to nitrocellulose membrane and probed with rabbit anti-cadherin (abcam, Cambridge, MA). The specific protein bands were visualized by a color metric substrate Opti-4CN (Bio-RAD).
C. Densitometry measurement of cadherin signals in B (TM vs. PM).
|Buffer A||25 ml||SM-005-A|
|Buffer B||10 ml||SM-005-B|
|Protein Extraction Filter Cartridges||50||P-001|
|Collection tubes with cap||50||P-002|
|Tissue dissociation beads||10 g||P-004|