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Minute™ 膜蛋白萃取套組 | Column Filter Invent Biotechnologies貨號SM-005

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http://www.inventbiotech.com/
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Minute™ 質膜蛋白萃取套組操作手冊

特點

Minute™  膜蛋白萃取套組 | Plasma Membrane Protein Isolation Kit 貨號SM-005  

sm-005-2

Minute™質膜(PM)蛋白分離試劑盒 (plasma membrane (PM) protein isolation kit);無洗滌劑和EDTA)的PM蛋白分離試劑盒;結合獨特的專利技術機制,細胞/組織只需通過Filter即完成質膜蛋白的萃取。在該過程中,細胞膜破裂不必使用傳統的均質化或超音波處理、不需要超速離心。處理量:1×106-50×106 cells/sample。時間:<45 min。Yield:1-3 mg/ml

The MinuteTM kit represents the next generation product for plasma membrane (PM) isolation and cell fractionation, offering unsurpassed convenience and consistency by eliminating uncontrollable variations associated with traditional homogenization and density gradient centrifugation including two phase partitioning.Compined-F

網站規格小圖_工作區域 1_工作區域 1

Minute™蛋白萃取套組,使用創新『Column Filter』技術,效果更佳,完成只需 一 分鐘蛋白萃取只需要vortex、transfer、spin 。Minute ™一系列蛋白質提取試劑盒;適用於多種物種(包括哺乳動物、海洋動物、昆蟲、植物和細菌)快速和容易地提取/分離蛋白質的優異工具 。與市場上的任何其他蛋白質提取試劑盒不同,優化的緩衝液系統,結合最新的專有濾筒技術(spin column);快速提高蛋白萃取的效能及方便性;用於總蛋白提取,細胞分離和膜蛋白分離,只需簡單四步驟即可完成蛋白質萃取。無須長時間等待、免除重複性的wash步驟。

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).

Item Amount Part #
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
Plastic rods 4 P-003
Tissue dissociation beads 10 g P-004

引用文獻  288+文獻資料請按此 

許多研究人員使用 Western blotting確認分離的膜蛋白之純度,一般常見的 “cytosolic markers” 不僅是exclusively cytosolic. 例如,肌動蛋白actin (1), GAPDH (2) and tubulin (3) 主要是胞質的蛋白; 但 也與質膜( plasma membranes)有關。 It’s not surprising to detect week signals of these marker proteins in PM preps in certain cell and tissue types. For more info please refer to following publications:

1. Gruenstein E., et al. (1975). Actin associated with membranes from 3T3 mouse fibroblast and Hela cells. Journal of cell Biology. 64:223-234.

2. Terrasse R., et al. (2012). Human and pneumococcal cell surface glyceraldehydes-3-phosphate dehydrogenase (GAPDH) proteins are both ligands of human C1q protein. J. Biol. Chem. 287:42620- 42633.

3. Wolff J. (2009). Plasma membrane tubulin. Biochemica et Biophysica Acta. (BBA)-Biomembranes 1788:1415-1433.

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invent

Fast Protein Extraction
Cat # Description Size
SD-001/SN-002 Total Protein Extraction Kit (for Animal Cultured Cells and Tissues) 50 preps
SN-006 Detergent-Free Protein Extraction Kit (for Animal Cultured Cells and Tissues) 50 preps
SD-008/SN-009 Total Protein Extraction Kit for Plant Tissues 50 preps
SN-010 Detergent-Free Plant Protein Extraction Kit 50 preps
MS-026 Total Protein Extraction Kit for Mass Spectrometry 50 preps
AT-022 Total Protein Extraction Kit for Adipose Tissues/Cultured Adipocytes 20 preps
SB-004 Bacterial Total Protein Extraction Kits 50 preps
YT-015 Total Protein Extraction Kit for Microbes with Thick Cell Walls 50 preps
YD-016 Detergent-Free Protein Extraction Kit for Microbes with Thick Cell Walls 50 preps
HD-021 Protein Extraction Kit for Hair and Nails 20 preps
FE-025 Protein Extraction Kit for Fixed and Embedded Tissues 20 preps

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Sub-Cellular Structure Fractionation/Isolation
Cat # Description Size
SC-003 Cytoplasmic & Nuclear Extraction Kits 50 Preps
SM-005 Plasma Membrane Protein Isolation Kit 50 preps
NE-013 Nuclear Envelope Protein Extraction Kit 50 preps
MP-007 Mitochondria Isolation Kit For Mammalian Cells And Tissues 50 preps
AF-023 Adipose Tissue Fractionation Kit 20 preps
NI-024 Detergent-free nuclei isolation kit 20 preps
YM-017 Yeast Mitochondria Enrichment Kit 50 preps
MM-018 Plant Microsomal Membrane Extraction Kit 50 preps
CP-011 Chloroplast Isolation Kit 50 preps

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Specialty Kits
Cat # Description Size
PN-019 Protein/Nucleic Acid Extraction Kit from Gels 50 preps
SC-012 Single Cell Isolation Kit 50 preps
HP-014 Histone/DNA Binding Protein Extraction Kit 50 preps

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Protein Analysis Reagents
Cat # Description Size
WA-001 HRP Substrate Kit 30 ml/30 ml
WA-002 Antibody Enhancer 30 ml
WA-003 Western Blot Stripping Solution 250 ml
WA-004 Ponceau S Stain 250 ml
WA-005 Peroxidase Suppressor 30 ml
WA-006 High Efficiency Protein Precipitation kit 30 ml/30 ml
WA-007 RBC Lysis Buffer 250 ml

引用文獻

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