ReadyPrep™ Protein Extraction Kit (Membrane II)

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ReadyPrep Protein Extraction Kit (Membrane II)

Overview

Fractionation of Proteins by Cellular Location

Differences in 2-D patterns using ReadyPrep signal (A), membrane I (B), and membrane II (C) kits. Mouse liver samples were extracted using the recommended protocols for each kit. Purified protein (~450 µg) was focused on 17 cm pH 3–10 nonlinear ReadyStrip IPG strips and run on 8–16% PROTEAN II precast gels for the second dimension. Overall spot patterns differ for A, B, and C even though all three kits isolate membrane proteins, indicating each kit isolates different types of membrane proteins.

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The ReadyPrep protein extraction kit (membrane II) applies sodium carbonate extraction methods (Fujiki et al. 1982, Molloy et al. 2000) to fractionate and isolate membrane proteins.

The ReadyPrep protein extraction kit (membrane II):

  • Isolates complex proteins that contain more than two transmembrane domains or those that are resistant to extraction by the ReadyPrep protein extraction (membrane I) kit
  • Requires ultracentrifugation
  • Includes a strongly chaotropic, IEF-compatible rehydration/sample buffer that contains the detergent ASB-14 to efficiently solubilize proteins

 

References
Fujiki Y et al. (1982). Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biology 93, 97–102.
Molloy MP et al. (2000). Proteomic analysis of the Escherichia coli outer membrane. Eur J Biochem 267, 2871–2881.

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ReadyPrep™ Protein Extraction Kit (Membrane II), 10 preps

1632084
10 preps, includes 2 x 5 ml 2-D Rehydration/Sample Buffer 1, 50 ml lysis buffer, 125 ml membrane protein concentrating reagent, 0.6 ml 200 mM tributylphosphine (TBP), 1 storage vial, instruction manual; for 10 extractions of 100–200 mg of cells or tissue

0 1632084 /templatedata/internet/product_SKU/data/LSR/Protein-Sample-Preparation/Protein-Fractionation/163-2084_0921120738728.xml 1632084 /default/main/bio-rad/WORKAREA/default 09/21/09 12:07 PM en_US juso9ms4 2018-02-01 00:53:48 true true internet/product_SKU juso9ms4 templatedata/internet/product_SKU/data/LSR/Protein-Sample-Preparation/Protein-Fractionation/163-2084_0921120738728.xml LSR 2019-05-04 16:23:43 /default/main/bio-rad/STAGING N /default/main/bio-rad en 12/31/37 11:59 PM 1556909733547 AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA,VN 2019-05-04 16:23:43 /default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Protein-Sample-Preparation/Protein-Fractionation/163-2084_0921120738728.xml templatedata/internet/product_SKU/data/LSR/Protein-Sample-Preparation/Protein-Fractionation/163-2084_0921120738728.xml 1632084-readyprep-protein-extraction-kit-membrane-ii-10-preps 1632084 ReadyPrep Protein Extraction Kit (Membrane II) ReadyPrep&trade; Protein Extraction Kit (Membrane II), 10 preps 10 preps, includes 2 x 5 ml 2-D Rehydration/Sample Buffer 1, 50 ml lysis buffer, 125 ml membrane protein concentrating reagent, 0.6 ml 200 mM tributylphosphine (TBP), 1 storage vial, instruction manual; for 10 extractions of 100–200 mg of cells or tissue 10 preps, protein preparation and fractionation kit for complex membrane proteins, Na carbonate extraction, includes strong detergent ASB-14 10 preps, includes 2 x 5 ml 2-D Rehydration/Sample Buffer 1, 50 ml lysis buffer, 125 ml membrane protein concentrating reagent, 0.6 ml 200 mM tributylphosphine (TBP), 1 storage vial, instruction manual; for 10 extractions of 100–200 mg of cells or tissue /webroot/web/images/lsr/products/protein_sample_preparation/sku_thumb/global/163-2084_tn.jpg /webroot/web/images/lsr/products/protein_sample_preparation/sku_view/global/163-2084_view.jpg /webroot/web/images/lsr/products/protein_sample_preparation/sku_feature/global/163-2084_feature.jpg ReadyPrep&trade; Protein Extraction Kit (Membrane II), 10 preps preview ,VN RepEx 163-2084 10 preps, protein preparation and fractionation kit for complex membrane proteins, Na carbonate extraction, includes strong detergent ASB-14 09/21/09 12:07 PM 12/31/37 11:59 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA,VN en LSR /LSR/Protein-Sample-Preparation/Protein-Fractionation N 0 <p>The ReadyPrep Protein Extraction Kit (Membrane II) is optimized for the isolation of proteins containing more than two transmembrane domains and proteins that are resistant to extraction by the ReadyPrep Membrane I Kit.</p> <ul> <li>Isolates complex proteins with 3 or more transmembrane domains and those resistant to extraction by the <a href="/en-us/sku/1632088-readyprep-protein-extraction-kit-membrane-i-50-preps?ID=1632088">ReadyPrep&trade; Protein Extraction (Membrane I) Kit</a></li> <li>Useful for a wide variety of biological samples, including animal tissue, animal cells, plant tissue, yeast, and bacteria</li> <li>Requires ultracentrifugation; entire procedure can be completed in less than 3 hours</li> <li>Includes a strongly chaotropic, IEF-compatible rehydration/sample buffer that contains the detergent ASB-14 to efficiently solubilize proteins</li> </ul> <p>&nbsp;<br /> The ReadyPrep Protein Extraction Kit (Membrane II) is based on the method of Fujiki et al. (1982) for the isolation of cellular membrane fractions after carbonate extraction, and the later adaption of the technique by Molloy et al. (2000). Incubation of tissue or cells with the high-pH Membrane Protein Concentrating Reagent strips weakly associated proteins away from membrane fragments. After ultracentrifugation and washing, the membrane pellet is solubilized using an IEF buffer or other buffer. The included 2-D Rehydration/Sample Buffer 1 contains the zwitterionic detergent ASB-14. The kit also contains the reducing agent tributylphosphine (TBP), which is more efficient than dithiothreitol (DTT) for keeping proteins reduced in 2-D PAGE applications.</p> <h2 class="descHeader">Kit Components</h2> <p>For 10 extractions of 100&ndash;200 mg of cells or tissue:</p> <ul> <li>2-D Rehydration/Sample Buffer 1, 2 vials, reconstituted total 10 ml</li> <li>Lysis Buffer, reconstituted 50 ml</li> <li>Membrane Protein Concentrating Reagent, reconstituted 125 ml</li> <li>TBP Reducing Agent, 1 ampoule; 0.6 ml of 200 mM tributylphosphine (TBP) in 1-methyl-2-pyrrolidinone sealed under nitrogen gas</li> <li>Empty vial for TBP storage </li> <li>Instruction manual</li> </ul> <p>Bio-Rad offers a range of ReadyPrep Kits for Fractionation by Cellular Location. These kits reduce sample complexity, increase detection and identification of low-abundance proteins, and simplify proteomic studies.</p> <table class="pd_table pd_gridlines" border="0"> <tbody> <tr> <td style="text-align: left;" width="200px" valign="top"><strong>ReadyPrep Kit for Fractionation by Cellular Location</strong></td> <td style="text-align: left;" valign="top"><strong>Proteins Targeted</strong></td> <td style="text-align: center;" valign="top"><strong>Catalog Number</strong></td> </tr> <tr class="pd_colorbackground"> <td valign="top">ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear)</td> <td style="text-align: left;" valign="top">Cytoplasmic and nuclear</td> <td style="text-align: center;" valign="top"><a href="/en-us/sku/1632089-readyprep-protein-extraction-kit-cytoplasmic-nuclear-50-preps?ID=1632089">1632089</a></td> </tr> <tr> <td valign="top">ReadyPrep Protein Extraction Kit (Membrane I)</td> <td style="text-align: left;" valign="top">Membrane-anchored, with one or two transmembrane domains, and hydrophobic</td> <td style="text-align: center;" valign="top"><a href="/en-us/sku/1632088-readyprep-protein-extraction-kit-membrane-i-50-preps?ID=1632088">1632088</a></td> </tr> <tr class="pd_colorbackground"> <td valign="top">ReadyPrep Protein Extraction Kit (Membrane II)</td> <td style="text-align: left;" valign="top">More than two transmembrane domains and/or proteins resistant to Membrane I Kit</td> <td style="text-align: center;" valign="top">1632084</td> </tr> <tr> <td valign="top">ReadyPrep Protein Extraction Kit (Signal)</td> <td style="text-align: left;" valign="top">Associated with lipid rafts</td> <td style="text-align: center;" valign="top"><a href="/en-us/sku/1632087-readyprep-protein-extraction-kit-signal-50-preps?ID=1632087-50-preps">1632087</a></td> </tr> </tbody> </table> <br /> <h2 class="descHeader">Related Products</h2> <ul> <li>ReadyPrep Kits for <a href="/en-us/category/fractionation-solubility">Fractionation by Solubility</a></li> <li>Aurum&trade; Ion Exchange Mini Kits for <a href="/en-us/category/fractionation-charge">Fractionation by Charge</a></li> <li>Rotofor<sup>&reg;</sup> Liquid-Phase IEF Cells for <a href="/en-us/category/fractionation-pi">Fractionation by pI</a></li> <li>Model 491 Prep Cell and Mini Prep Cell for <a href="/en-us/category/fractionation-solubility">Fractionation by Solubility </a></li> <li><a href="/en-us/product/readyprep-2-d-starter-kit">ReadyPrep&trade; 2-D Starter Kit</a></li> <li><a href="/en-us/product/protean-i12-ief-system">PROTEAN<sup>&reg;</sup> i12&trade; IEF System</a></li> <li><a href="/en-us/product/readystrip-ipg-strips">ReadyStrip&trade; IPG Strips</a></li> <li>ReadyPrep Reduction-Alkylation Kit (<a href="/en-us/sku/1632090-readyprep-reduction-alkylation-kit-100-preps?ID=1632090">1632090</a>)</li> <li><a href="/en-us/product/powerpac-hv-high-voltage-power-supply">PowerPac&trade; HV High-Voltage Power Supply</a></li> </ul> <h2 class="descHeader">Related Categories</h2> <ul> <li><a href="/en-us/category/protein-gels">Protein Gels</a></li> <li><a href="/en-us/category/electrophoresis-chambers">Electrophoresis Chambers</a></li> <li><a href="/en-us/category/2-d-electrophoresis?ID=6916a228-0f95-4b26-a37a-5e4afe76add7-buffers-reagents">2-D Electrophoresis Buffers and Reagents</a></li> <li><a href="/en-us/product/ampholytes">Ampholytes</a></li> <li><a href="/en-us/category/tube-gel-ief-systems">Tube Gel IEF Systems</a></li> <li><a href="/en-us/category/imaging">Gel Imaging Systems</a></li> <li><a href="/en-us/product/2-d-electrophoresis-workflow">2-D Electrophoresis Workflow</a></li> </ul> <h2 class="descHeader">Supporting Documentation</h2> <ul> <li>ReadyPrep Protein Extraction Kit (Membrane II) <a href="/webroot/web/pdf/lsr/literature/4110157A.pdf" target="_blank">Instruction Manual</a></li> <li>Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion <a href="/webroot/web/pdf/lsr/literature/Bulletin_3096.pdf" target="_blank">Brochure</a></li> <li>ReadyStrip IPG Strip <a href="/webroot/web/pdf/lsr/literature/4006166H.pdf" target="_blank">Instruction Manual</a></li> <li>PROTEAN i12 IEF System <a href="/webroot/web/pdf/lsr/literature/Bulletin_6097A.pdf" target="_blank">Brochure</a></li> <li>Host Cell Protein (HCP) Workflow <a href="/webroot/web/pdf/lsr/literature/Bulletin_6493.pdf" target="_blank">Quick Guide</a></li> </ul> <h2 class="descHeader">References</h2> <p>Fujiki Y et al. (1982). Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biology 93, 97&ndash;102. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/7068762">7068762</a><br /> Molloy MP et al. (2000). Proteomic analysis of the Escherichia coli outer membrane. Eur J Biochem 267, 2871&ndash;2881. PMID: <a href="http://www.ncbi.nlm.nih.gov/pubmed/10806384">10806384</a></p> /webroot/web/html/lsr/products/SKUs/autopopulated 1632084.html /webroot/web/html/lsr/products/SKUs/autopopulated/1632084.html false $306.00 false $ null true false true true false USD true true true /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1632084&quantity= false false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1632084&CartQuote=QT&quantity= 1432 true true false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1632084&AddToDefaultWishList=true&quantity= 41106500 306.00 1632084
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