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Minute™ Total Protein Extraction Kit for Bone Tissue

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Minute™ Total Protein Extraction Kit for Bone Tissue (50 Preps)

Cat #: SA-02-BT

Description

Bone tissues are commonly used for research. There are two types of bone tissues: compact and spongy. Cells in bone tissues are tightly packed and, because of their unique structures, it is very difficult to efficiently extract total protein from them. Traditional solution-based protein extraction methods, such as RIPA, are inefficient and the protein yield is very low. The profile of extracted protein is also incomplete with solution-based methods. This kit provides a highly efficient method for extracting proteins from human or animal bone tissue by a combination of mechanical extraction and chemical lysis of cells in tissues. The kit features a simple and fast single-tube protocol and optimized buffers for bone tissue. Researchers have the option to choose either a denaturing or native cell lysis buffer, which is specifically tailored for bone tissue. The whole procedure takes less than 10 minutes to complete and the protein yield is in the range of 0.5-2 mg/ml depending upon type of bones. The materials provided are sufficient for 50 extractions.

Shipping and storage: This kit is shipped and stored at room temperature.

Kit includes:

Items

Quantity

Denaturing Buffer

25 ml

Native Buffer

25 ml

Protein Extraction Powder

5g

Plastic Rods

2 units

Filter Cartridges  

50 units

Collection Tubes

50 units

References (10):

1. Ren, H., Wang, Z., Xu, J., Chen, J., & Lan, J. (2020). The Impact of Frizzled-9 on Dental Implant Osseointegration in Hyperlipidemic Rats. Journal of Hard Tissue Biology, 29(1), 37-44.

2. Pedersen, K. B., Osborn, M. L., Robertson, A. C., Williams, A. E., Watt, J., Denys, A., … & Ronis, M. J. (2020). Chronic Ethanol Feeding in Mice Decreases Expression of Genes for Major Structural Bone Proteins in a Nox4-Independent Manner. Journal of Pharmacology and Experimental Therapeutics, 373(3), 337-346.

3.  Liu, H. D., Ren, M. X., Li, Y., Zhang, R. T., Ma, N. F., Li, T. L., … & Yang, M. (2022). Melatonin alleviates hydrogen peroxide induced oxidative damage in MC3T3-E1 cells and promotes osteogenesis by activating SIRT1. Free Radical Research, (just-accepted), 1-19.

4.  Jiang, C., Lin, Y., Shan, H., Xia, W., Pan, C., Wang, N., … & Yu, X. (2022). miR-146a Protects against Staphylococcus aureus-Induced Osteomyelitis by Regulating Inflammation and Osteogenesis. ACS Infectious Diseases.

5. Song, D., Ha, C. Z., Xu, Q., & Hu, Y. H. (2022). mRNA expression profiling of the cancellous bone in patients with idiopathic osteonecrosis of the femoral head by whole-transcriptome sequencing. Medicine, 101(35), e30213.

6. Deng, J., Cohen, D. J., Sabalewski, E. L., Van Duyn, C., Wilson, D. S., Schwartz, Z., & Boyan, B. D. (2022). Semaphorin 3A Delivered by a Rapidly Polymerizing Click Hydrogel Overcomes Impaired Implant Osseointegration in a Rat Type 2 Diabetes Model. Acta Biomaterialia.

7.  He, H., Zhang, Y., Sun, Y., Zhang, Y., Xu, J., Yang, Y., & Chen, J. (2022). Folic Acid Attenuates High-Fat Diet-Induced Osteoporosis Through the AMPK Signaling Pathway. Frontiers in cell and developmental biology, 9, 3661.

8.  Choi, R. Y., Kim, I. W., Ji, M., Paik, M. J., Ban, E. J., Lee, J. H., … & Seo, M. (2023). Protaetia brevitarsis seulensis larvae ethanol extract inhibits RANKL-stimulated osteoclastogenesis and ameliorates bone loss in ovariectomized mice. Biomedicine & Pharmacotherapy, 165, 115112.

9.  Liu, Z., Yao, X., Jiang, W., Zhou, Z., & Yang, M. (2023). Sodium butyrate enhances titanium nail osseointegration in ovariectomized rats by inhibiting the PKCα/NOX4/ROS/NF-κB pathways. Journal of Orthopaedic Surgery and Research, 18(1), 1-14.

10.  Zhang, C. X., & Man, Q. W. (2024). Proteomics study of bone tissue around ameloblastoma and the potential mechanism of CD36 in bone remodelling. British Journal of Oral and Maxillofacial Surgery.

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Minute™ Total Protein Extraction Kit for Skin Tissue

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Minute™ Total Protein Extraction Kit for Skin Tissue (50 Preps)

Cat #: SA-01-SK

Description

kin tissue is made up of epidermis, dermis and subcutaneous fat. Because of its unique structure, skin tissue is notoriously difficult to homogenize. It is also very difficult to lyse the cells in the tissue for total protein extraction. Traditional solution-based protein extraction methods, such as RIPA, are inefficient and the yield is very low. The profile of extracted protein is also incomplete with solution-based methods. This kit provides a highly efficient method for total protein extraction from human or animal skin tissues by using a combination of mechanical extraction and chemical lysis of skin tissues. The kit features a simple and fast single-tube protocol and optimized buffers for skin tissues. The researchers have the option to choose either a denaturing or native cell lysis buffer, which is specifically tailored for skin tissue. The whole procedure takes less than 10 minutes to complete and the protein yield is in the range of 1-5 mg/ml. The materials provided are sufficient for 50 extractions.

Shipping and storage: This kit is shipped and stored at room temperature.

 

Kit includes:

Items

Quantity

Denaturing Buffer

25 ml

Native Buffer

25 ml

Protein Extraction Powder

5g

Plastic Rods

2 units

Filter Cartridges  

50 units

Collection Tubes50 units

References (11)

  1. Zhang, C., Li, Y., Qin, J., Yu, C., Ma, G., Chen, H., & Xu, X. (2021). TMT-Based Quantitative Proteomic Analysis Reveals the Effect of Bone Marrow Derived Mesenchymal Stem Cell on Hair Follicle Regeneration. Frontiers in Pharmacology, 12.
  2. Wang, T., Zhu, G., Qin, L., Wang, Q., She, C., Xu, D., … & Li, S. T. (2021). Kininogen–Nitric Oxide Signaling at Nearby Nonexcited Acupoints after Long-Term Stimulation. JID Innovations, 1(3), 100038.
  3. Yin, S., Song, R., Ma, J., Liu, C., Wu, Z., Cao, G., … & Wang, Y. (2022). Receptor activity‐modifying protein 1 regulates mouse skin fibroblast proliferation via the Gαi3-PKA-CREB-YAP axis. Cell Communication and Signaling, 20(1), 1-18.
  4. Wang, Y., Huang, J., & Jin, H. (2022). Reduction of Methyltransferase-like 3-Mediated RNA N6-Methyladenosine Exacerbates the Development of Psoriasis Vulgaris in Imiquimod-Induced Psoriasis-like Mouse Model. International Journal of Molecular Sciences, 23(20), 12672.
  5. Zhu, W., Dong, Y., Xu, P., Pan, Q., Jia, K., Jin, P., … & Che, B. (2022). A composite hydrogel containing resveratrol-laden nanoparticles and platelet-derived extracellular vesicles promotes wound healing in diabetic mice. Acta Biomaterialia.
  6. Yuan, X., Tang, B., Chen, Y., Zhou, L., Deng, J., Han, L., … & Wang, Y. (2023). Celastrol inhibits store operated calcium entry and suppresses psoriasis. Frontiers in Pharmacology, 14.
  7. Zhou, H., Gan, Y., Li, Y., Chen, X., Guo, Y., & Wang, R. (2023). Degradation of Rat Sarcoma Proteins Targeting the Post-Translational Prenyl Modifications via Cascade Azidation/Fluorination and Click Reaction. Journal of Medicinal Chemistry.
  8. Wang, J. Study on the Regulatory Mechanism of Jia Wei Guo Min Decoction On Degranulation of Mast Cell in Ad Mice Based on His/Par-2 Pathway.
  9. Huang, J. L., Xu, Y. H., Yang, X. W., Wang, J., Zhu, Y., & Wu, X. B. (2023). Jiawei guomin decoction regulates the degranulation of mast cells in atopic dermatitis mice via the HIS/PAR-2 pathway. Journal of Ethnopharmacology, 117485.
  10. Wu, Z., Liu, C., Yin, S., Ma, J., Sun, R., Cao, G., … & Wang, Y. (2024). P75NTR regulates autophagy through the YAP-mTOR pathway to increase the proliferation of interfollicular epidermal cells and promote wound healing in diabetic mice. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 167012.
  11.  Xu, X. L., Wu, S. J., Qi, S. Y., Chen, M. M., Liu, Z. M., Zhang, R., … & Lian, Z. X. (2024). Increasing GSH-Px Activity and Activating Wnt Pathway Promote Fine Wool Growth in FGF5-Edited Sheep. Cells, 13(11), 985.
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Minute™ Protein Extraction Kit for Fixed and Embedded Tissues

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Minute™ Protein Extraction Kit for Fixed and Embedded Tissues (20 Preps)

Cat #: FE-025

Description

Formalin-fixed and paraffin-embedded (FFPE) tissues are widely available and serve as a potentially rich source for biomedical research. However, protein extraction from FFPE tissue is very challenging due to formalin-mediated molecular cross-linking. Traditional methods for protein extraction from FFPE tissues involve repeated deparaffinization and rehydration using organic solvents followed by protein extraction at a high temperature with or without an optional sonication step. These methods can be effective but are tedious and time-consuming (3-4 hours). Minute™ Protein Extraction Kit for Fixed and Embedded Tissues provides a simple and rapid way to extract protein from FFPE tissue without organic solvent deparaffinization. The whole procedure can be completed in less than one hour with a protein yield of 2-3 mg/ml.

Kit includes:

Items

Quantity

Buffer A

30 ml

Buffer B

10 ml

1.5 ml Microfuge Tubes

20 units

Pestles

2 units

Protein Extraction Powder2 grams

 

References (3)

1. Gnanaguru, G., Wagschal, A., Oh, J., Saez-Torres, K. L., Li, T., Temel, R. E., … & D’Amore, P. A. (2021). Targeting of miR-33 ameliorates phenotypes linked to age-related macular degeneration. Molecular Therapy.

2. Obi, E. N., Tellock, D. A., Thomas, G. J., & Veenstra, T. D. (2023). Biomarker Analysis of Formalin-Fixed Paraffin-Embedded Clinical Tissues Using Proteomics. Biomolecules, 13(1), 96.

3.  Moriyama, R., Nakamura, S., Mitsui, I., Sugiyama, M., Fukui, H., Fukui, H., … & Suzuki, J. (2023). Expression of SARS-CoV-2 entry molecules ACE2, NRP1, TMPRSS2, and FURIN in the reproductive tissues of male macaques.

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Protein Extraction Kit for Hair and Nails

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Protein Extraction Kit for Hair and Nails (20 Preps)

Cat #: HD-021

Description

Proteins account for about 80% of keratinized animal tissues such as hair, nail, horn and wool. These tissues contain mainly hard keratins made of several distinct types of protein with a molecular weight ranging from 10 to 135 kDa. The extraction buffer usually contains a high concentration of urea, thiourea, guanidine hydrochloride or a combination of these strong denaturants. Protein extraction with high concentration denaturants is relatively effective, but because of larger extraction buffer/tissue ratio, the concentration of extracted protein is usually low. Extracted proteins need to be concentrated prior to further analysis. The presence of high-concentration denaturants may also interfere with downstream application. HD-021 is designed to effectively extract proteins from keratinized animal tissues without using strong denaturants. The extracted protein can be directly loaded onto SDS-PAGE without concentration. The extraction buffer of this kit contains 0.5% SDS and other chemicals. Extracted protein concentration is 1-2 mg/ml.

Kit includes:

Items

Quantity

Buffer A

15 ml

Buffer B

1.5 ml

Filter Cartridges

20 units

Collection Tubes with Cap

20 units



References (7)
  1. Lee, S. H., Baldina, E., Lee, E., & Youm, Y. (2021). Social connectedness and hair cortisol in community-dwelling older adults. Comprehensive Psychoneuroendocrinology, 100053.
  2. Marsden, A. J., Riley, D. R., Birkett, S., Rodriguez-Barucg, Q., Guinn, B. A., Carroll, S., … & Beltran-Alvarez, P. (2021). Love is in the hair: arginine methylation of human hair proteins as novel cardiovascular biomarkers. Amino Acids, 1-10.
  3. Kim, W. J., Park, K. M., Park, J. T., Seo, E., An, S. K., Park, H. Y., & Lee, E. (2021). Sex-specific association of hair cortisol concentration with stress-related psychological factors in healthy young adults. Biology of Sex Differences, 12(1), 1-7.
  4. Kim, W. J., Park, K. M., Park, J. T., Seo, E., Bang, M., An, S. K., … & Lee, E. (2021). Effect of Childhood Trauma on the Association Between Stress-Related Psychological Factors and Hair Cortisol Level in Young Adults. Psychiatry Investigation, 18(11), 1131.
  5. Lee, S., Park, J. T., Bang, M., An, S. K., Namkoong, K., Park, H. Y., & Lee, E. (2022). Theory of mind and hair cortisol in healthy young adults: the moderating effects of childhood trauma. European Journal of Psychotraumatology, 13(2), 2116826.
  6. Fuse, N., Morita, S., & Matsue, Y. (2023). Age‐related hair denaturation related to protein carbonyls. International Journal of Cosmetic Science.
  7. Fuse, N., Morita, S., & Matsue, Y. (2023). Age‐related hair denaturation related to protein carbonyls. International Journal of Cosmetic Science.
  8. Li, Z., Kong, W., Park, H. Y., Koo, S. J., Bang, M., Park, J. T., … & An, S. K. (2024). Association of hair cortisol concentration with brain‐derived neurotrophic factor gene methylation: The role of sex as a moderator. Stress and Health, e3401.
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Minute™ Detergent-Free Protein Extraction Kit for Microbes with Thick Cell Walls

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Minute™ Detergent-Free Protein Extraction Kit for Microbes with Thick Cell Walls (50 Preps)

Cat #: YD-016

Description

Protein extraction from cell-wall-containing microbes is a common procedure. The methods for protein extraction are usually harsh, tedious and unreliable. YD-016 provides a detergent-free, rapid and gentle way for extracting proteins from microbes with thick and strong cell walls. These microbes include yeast, filamentous fungus, gram-positive and -negative bacteria, insect eggs, microalgae and more. This kit contains optimized detergent-free and EDTA-free protein extraction buffers. The whole procedure takes less than 10 minutes and the protein yield is in the range of 2-4 mg/ml with unbiased protein representation. This kit contains a single-tube protocol and works well for both log-phase and stationary-phase microbes. The materials provided are sufficient for 50 extractions.

Kit includes:

Items

Quantity

Buffer A

20 ml

Buffer B

20 ml

Protein Extraction Powder

5 grams

1.5 ml Microfuge Tube

50 units

Pestles

2 units

 



References (1)
  1. Peng, G., Hou, X., Zhang, W., Song, M., Yin, M., Wang, J., … & Li, X. (2018). Alkyl rhamnosides, a series of amphiphilic materials exerting broad-spectrum anti-biofilm activity against pathogenic bacteria via multiple mechanisms. Artificial cells, nanomedicine, and biotechnology, 1-16.
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Minute™ Bacterial Total Protein Extraction Kit

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Minute™ Bacterial Total Protein Extraction Kit (50 Preps)

Cat #: SB-004

Description

Minute™ Bacterial Total Protein Extraction Kit is composed of optimized denaturing cell lysis buffers and protein extraction filter cartridges with 2.0 ml collection tubes. The kit is designed to quickly extract denatured proteins from bacteria. It contains sufficient materials for the extraction of total proteins from a 100 ml E. coli culture.

Kit includes:

Items

Quantity

Bacterial Lysis Buffer A

25 ml

Buffer B

2.5 ml

Protein Extraction Filter Cartridges

50 units

Collection Tubes with Caps

50 units

 



References (2)
  1. http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_Newsletter_2013-09_Resource_Notes_Fall.pdf#page=9
  2. Yonezawa, H., Osaki, T., Fukutomi, T., Hanawa, T., Kurata, S., Zaman, C., … & Kamiya, S. (2017). Diversification of the AlpB outer membrane protein of Helicobacter pylori affects biofilm formation and cellular adhesion. Journal of Bacteriology, 199(6), e00729-16.
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Minute™ Total Protein Extraction Kit for Adipose Tissues/Cultured Adipocytes

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Minute™ Total Protein Extraction Kit for Adipose Tissues/Cultured Adipocytes (20 Preps)

Cat #: AT-022

Description

Adipose tissue, especially white adipose tissue (WAT), has been recognized as an essential endocrine and inflammation organ in addition to its energy storage function. Analyses of proteins from adipose tissues are increasingly critical for understanding many physiological/pathological conditions. However, isolation of WAT and brown adipose tissue (BAT) is technically challenging due to their high lipid and low protein contents. The water-oil emulsion present in a biological sample is notoriously difficult to separate. We have developed a novel technology to address this issue. A porous filter with unique surface property and pre-defined pore size and thickness coupling with a specially formulated detergent-free extraction buffer is employed to rapidly and effectively separate water-oil emulsion derived from adipose tissue homogenate. The extraction buffer has a lower freezing point than oil in adipose tissues, and the aqueous phase can be quickly separated from the oil phase by passing the tissue homogenate through the filter. The total proteins isolated are the unbiased representation of cellular proteins in the tissue. The extracted proteins concentration is very high (2-3 mg/ml) compared to traditional methods.

 Kit includes:

Items

Quantity

Buffer A (Extraction Buffer)

15 ml

Buffer B (10 X Denaturing Buffer)

1.5 ml

Buffer C (10 X Non-Denaturing Buffer)

1.5 ml

1.5 ml Microfuge Tubes

20 units

Pestles for 1.5 ml Tubes

2 units

Filter Cartridge with Collection Tubes

20 units

Protein Extraction Powder

2 grams

A. SDS-PAGE (10%) profiles of total protein extracted from different adipose tissues. Lane 1, porcine WAT; Lane 2, chicken BAT; Lane 3, rat WAT.

B. Western blottings of extracted proteins from rat WAT. Proteins were separated in 8-16% gradient SDS-PAGE and probed with following cellular protein marker antibodies:

Anti-Na/K ATPase alpha1, a plasma membrane marker (Upstate, clone 464.6), anti-lamin B1, a nuclear envelope marker (ab16048, abcam Cambridge, MA), anti-ubiquinol-cytochrome C reductase core protein (abcam, ab 96333) and GAPDH, a cytosolic marker (Sigma). The specific protein bands were visualized by a substrate Opti-4CN (Bio-RAD).

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Minute™ Total Protein Extraction Kit for Mass Spectrometry

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Minute™ Total Protein Extraction Kit for Mass Spectrometry (50 Preps)

Cat #: MS-026

Description

Mass spectrometry has become a popular method for analyzing and characterizing proteins. Traditional total protein extraction methods, such as RIPA-based reagents, can alter endogenous baselines due to nonsystematic protein loss. Some chemicals in RIPA may not be compatible with mass spectrometry and other downstream applications. This kit extracts total proteins quickly using a specially formulated lysis buffer and proprietary spin column. The extracted proteins are compatible with trypsin digestion followed by MS analysis, iTRAQ, biotin labeling, etc. Due to the use of filter cartridges, the extraction volume can be as low as 20 µl – a helpful feature when starting material is a limiting factor. The typical yield is 2-8 mg/ml.

Kit includes:

Items

Quantity

Lysis Buffer

25 ml

Protein Extraction Filter Cartridges

50 units

Collection Tubes with Caps

50 units

Plastic Rods

2 units

References (1)
  1. Chen, Y., Fang, Z., Zhou, J., Qin, H., & Ye, M. (2021). Mirror-Cutting-Based Digestion Strategy Enables the In-Depth and Accuracy Characterization of N-Linked Protein Glycosylation. Journal of Proteome Research.
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Minute™ Cytoplasmic and Nuclear Extraction Kits for Cells

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Minute™ Cytoplasmic and Nuclear Extraction Kits for Cells (50 Preps)

Cat #: SC-003

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Description

Minute™ Cytoplasmic and Nuclear Extraction Kit for Cells comprises optimized cytoplasmic extraction buffer, nuclear protein extraction buffer, and protein extraction filter cartridges with 2.0 ml collection tubes. The kit is designed to quickly separate native cytosol and nuclear proteins from mammalian cells and protoplasts of plants, bacteria, yeast, and fungus. Separation of cytoplasmic and nuclear proteins can be accomplished in less than 15 minutes using the protein extraction filter cartridges.

Kit includes:

Items

Quantity

Cytoplasmic Extraction Buffer

25 ml

Nuclear Extraction Buffer

25 ml

Protein Extraction Filter Cartridges

50 units

Collection Tubes with Caps

50 units

References (262)
  1. Kalabus, JL. et al. (2012). MicroRNAs differentially regulate carbonyl reductase 1 (CBR1) gene expression dependent on the allele status of common polymorphic variant rs9024. PLOS ONE. 7(11):1-9.
  2. Lombrafia AQ. et al (2014). Interindividual variability in the cardiac expression of anthracycline reductases in donors with and without down syndrome.  Pharm Res. 31:1644-1655.
  3. Hou, N., et al. (2014). Vitamin A deficiency impairs spatial learning and memory: The mechanism of abnormal CBP-dependent histone acetylation regulated by retinoic acid receptor alpha. Mol Neurobiol.  DOI 10.1007/s12035-014-8741-6.
  4. You, Y., et al. (2014). Latexin sensitizes leukemogenic cells to gamma-irradiation-induced cell-cycle arrest and cell death through Rps3. Cell Death and Disease. (2014)5, e1493; doi: 10.1038/cddis. 2014.443.
  5. Tanaka M et al. (2016). Ex vivo effects of naphthoquinones allergen-sensitized mononuclear cells in mice. International Journal of Immunopathology and Pharmacology. DOI: 10.1177/0394632016632221
  6. Liu W. et al. (2016) Electroacupunctre improves motor impairment via inhibition of microglia-mediated neuroinflammation in the sensorimotor cortex after ischemic stroke. Life Sciences. doi:10.1016/j.lfs.2016.01.045
  7. Xu, Yang, et al. (2016), RIP3 induces ischemic neuronal DNA degradation and programmed necrosis in rat via AIF.” Scientific Reports 6:29362.
  8. Shi, Le, et al. (2016). “Evodiamine exerts anti-tumor effects against hepatocellular carcinoma through inhibiting β-catenin-mediated angiogenesis.” Tumor Biology: 1-13.9
  9. Huang, C. W., Feng, W., Peh, M. T., Peh, K., Dymock, B. W., & Moore, P. K. (2016). A novel slow-releasing hydrogen sulfide donor, FW1256, exerts anti-inflammatory effects in mouse macrophages and in vivo. Pharmacological Research.
  10. Matsuoka, Y., Nakayama, H., Yoshida, R., Hirosue, A., Nagata, M., Tanaka, T., … & Shinriki, S. (2016). IL-6 controls resistance to radiation by suppressing oxidative stress via the Nrf2-antioxidant pathway in oral squamous cell carcinoma. British Journal of Cancer.
  11. Meng, C., Cui, X., Qi, S., Zhang, J., Kang, J., & Zhou, H. (2016). Lung inflation with hydrogen sulfide during the warm ischemia phase ameliorates injury in rat donor lungs via metabolic inhibition after cardiac death. Surgery.
  12. Liu, Y., Peng, J., Sun, T., Li, N., Zhang, L., Ren, J., … & Ding, Y. (2017). Epithelial EZH2 serves as an epigenetic determinant in experimental colitis by inhibiting TNFα-mediated inflammation and apoptosis. Proceedings of the National Academy of Sciences, 201700909.
  13. Chen, Y., Hu, X., Guo, C., Yu, Y., & Lu, H. (2017). Epe1 contributes to activation of AMPK by promoting phosphorylation of AMPK alpha subunit, Ssp2. Scientific Reports, 7.
  14. Hu, H., Zhu, W., Qin, J., Chen, M., Gong, L., Li, L., … & Ye, D. (2016). Acetylation of PGK1 Promotes Liver Cancer Cell Proliferation and Tumorigenesis. Hepatology
  15. Hou, Y., Wang, Y., He, Q., Li, L., Xie, H., Zhao, Y., & Zhao, J. (2017). Nrf2 Inhibits NLRP3 Inflammasome Activation Through Regulating Trx1/TXNIP Complex in Cerebral Ischemia Reperfusion Injury. Behavioural Brain Research.
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Minute™ Plant Plasma Membrane/Protein Isolation Kit

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Minute™ Plant Plasma Membrane/Protein Isolation Kit (50 Preps)

Cat #: SM-005-P

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Description

Plasma membrane (PM) protein accounts for a small fraction of total cellular protein in plants but performs a very critical role in plant physiology. Isolation and purification of PM protein from plant tissues have been traditionally done by sucrose density ultracentrifugation and aqueous two-phase partitioning. These relatively effective methods require ultracentrifugation and a large amount of starting material. The procedures are usually tedious and time-consuming. To overcome the shortcomings, we have developed this PM isolation kit. Plant tissues are first sensitized by buffer A, homogenized and then passed through a specialized filter cartridge that allows homogenates to pass through with a zigzag path. The cell membranes are ruptured into a range of predefined sizes during the process. Native plasma membranes are separated from a mixture of unruptured cells, nuclei, cytosol and organelles by subsequent differential centrifugation and density centrifugation without using ultracentrifugation. Due to the use of the same amount of starting material, defined centrifugal force and predefined duration in every experiment, the result is much more consistent with a high degree of PM protein enrichment. The procedure can be completed in about 1 hour.

Kit includes:

Items

Quantity

Buffer A

25 ml

Buffer B

10 ml

Protein Extraction Filter Cartridges

50 units

Collection Tubes with Caps

50 units

Plastic Rods

2 units

Tissue Dissociation Beads

10 grams

 



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