Introduction to Protein Sample Preparation

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en-us LUSP108WI Protein Sample Preparation Introduction to Protein Sample Preparation /webroot/web/html/lsr/solutions/technologies/protein_electrophoresis <p>Protein sample preparation generally involves some or all of the following steps: cell disruption, extraction, removal of interfering compounds, changes of the physio/chemical properties and concentration of the sample. The quality of sample preparation can greatly affect the quality of the data that are generated. General guidelines and some of the most common methods for protein sample preparation are provided in this section. Protocols for sample preparation from mammalian tissue, plant leaves, microbial cultures, and for various buffer formulations are provided at the bottom of the page in the <a href="#related_content">protocols</a> tab.</p> <p><strong>Related Sections</strong>: <a href="/evportal/destination/solutions?catID=LUSP8GD57">Protein Electrophoresis Reagents Selection and Preparation</a></p> Protein Sample Preparation Workflow <p><img class="stroke" src="http://www.bio-rad.com/webroot/web/images/1x1grey.gif" alt="" /></p> <p><img usemap="#Map" src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/protein_electrophoresis/technology_detail/pet13_img1.jpg" border="0" alt="" width="473" height="691" /> <map name="Map"> <area shape="rect" coords="5,4,475,143" href="/evportal/destination/solutions?catID=LUSP2D1FX"></area> <area shape="rect" coords="5,142,472,280" href="/evportal/destination/solutions?catID=LUSP3KFD4"></area> <area shape="rect" coords="4,278,475,442" href="/evportal/destination/solutions?catID=LUSP4LJSZ"></area> <area shape="rect" coords="4,441,476,580" href="/evportal/destination/solutions?catID=LUSP5JKAJ"></area> <area shape="rect" coords="5,579,474,692" href="#"></area> </map> </p> <p class="caption"><strong>Protein sample preparation workflow.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> General Considerations <p>Due to the great diversity of protein sample types and sources, no single sample preparation method works with all proteins; for any sample, the optimum procedure must be determined empirically. However, the following general sample preparation guidelines should be kept in mind to avoid a number of pitfalls during sample preparation for protein electrophoresis (Posch et al. 2006).</p> <ul> <li>Keep the sample preparation workflow as simple as possible (increasing the number of sample handling steps may increase variability)</li> <li>With cell or tissue lysates, include protease inhibitors to minimize artifacts generated by proteolysis; protease inhibitors are generally not required for samples like serum or plasma</li> <li>Solubilize proteins in a buffer that is compatible with the corresponding electrophoresis technique</li> <li>Determine the amount of total protein in each sample using a protein assay that is compatible with chemicals in your samples</li> <li>Use protein extracts immediately or aliquot them into appropriately sized batches and store them at &ndash;70&deg;C to avoid freeze-thaw cycles</li> </ul> <div class="top"><a href="#helptop">Back to Top</a></div> References <p>Posch A et al. (2006). Tools for sample preparation and prefractionation in two-dimensional gel electrophoresis. In Separation methods in Proteomics, Smejkal GB ed. (Boca Raton: CRC Press), pp 107&ndash;133.</p> <div class="top"><a href="#helptop">Back to Top</a></div> <p><a name="related_content"></a></p> Protocols <table id="carttablealigned" class="literature_table" style="height: auto; width: 583px;" border="0" cellspacing="0" cellpadding="0"> <tbody> <tr> <td width="100">6194</td> <td width="350">Protein Sample Generation General Tips</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6194.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6195</td> <td width="350">Human Cells Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6195.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6196</td> <td width="350">Mammalian Tissue Protcol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6196.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6197</td> <td width="350">Plant Leaves Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6197.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6198</td> <td width="350">Microbial Cultures Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6198.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6199</td> <td width="350">Buffer Formulations Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6199.pdf" target="_blank"><span>Click to download</span></a></td> </tr> </tbody> </table> 3096 Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion Brochure, Rev B 3096 /webroot/web/pdf/lsr/literature/Bulletin_3096.pdf Literature PDF Brochures_and_Specifications /webroot/web/images/general/icons/icon_pdf.gif Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion Brochure No Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion Brochure, Rev B 3096 bio-spin, dynamic range reduction, sample prep, preparation, prep cell, rotofor, enrichment of low-abundance, isoelectric focusing, lit3096, membrane i, micro bio spin columns, model 491, proteins, truview cuvettes, cell lysis, expression proteomics, mini, prep, proteominer kits, quick start bradford assay, ready prep, readyprep, 2-d electrophoresis, affi-gel blue, albumin, aurum, chemical reagents, ief, microrotofor, protean ii, rc dc, smartspec plus, spectrophotometer, 165-2435, 1652435 6040 /templatedata/internet/documentation/data/LSR/Literature/6040.xml Electrophoresis Guide, Interactive PDF, Rev C 6040 /webroot/web/pdf/lsr/literature/Bulletin_6040.pdf Literature PDF Manuals_and_Quick_Guides /webroot/web/images/general/icons/icon_pdf.gif No Electrophoresis Guide, Interactive PDF, Rev C 6040 Bulletin 2895, 6040, overview and principle, theory and product selection, one dimensional (1-D) protein separation, blue native polyacrylamide, SDS-PAGE, sodium dodecyl sulfate, sodium dodecyl sulphate, polymerization, precast, handcast resolving, stacking single-percentage gel, Laemmli (Tris-HCl), Bis-Tris, Tris-acetate, Tris-Tricine premade running and sample buffers, formulations/components, reagents, salts, isoelectric focusing (IEF), zymogram, gel casting chambers, gradient formers, sample preparation protocols, cell lysis, disruption, homogenization, protein solubilization, human suspension/monolayer cultured cells, mammalian tissue, plant leaves, microbial cultures, chromatography protein fractions, reducing, chaotropic agents, detergents, sample loading and quantitation spectrophotometric assays, recombinant and natural protein standards, prestained or unstained molecular weight markers or ladders, selecting power supplies, joule heating, separations under constant voltage, current, power, detection and analysis, total and specific protein staining, Coomassie Blue stain, fluorescent gel and silver stains, stainers, imagers, imaging systems and software, protein molecular weight (size) estimation, quantification, downstream applications, western blotting, gel drying, electroelution, spot excision (cutting), troubleshooting tips Life Science Research/Products/Protein Sample Preparation/Protein Extraction/MicroRotofor Cell Lysis Kits ->MT::5e97c5be-3707-4c08-ae55-25e883f2ab99##Life Science Research/Products/Protein Sample Preparation/Protein Extraction/ReadyPrep Mini Grinders ->MT::0e0a3350-ceb8-4c52-9346-8f48ecee6e5a##Life Science Research/Products/Sample Quantitation/Protein Assay Kits and Cuvettes/<em>RC DC</em> Protein Assay ->MT::f58fdf1d-5d0b-4f31-93e2-f812bd7e7eab##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Electrophoresis Buffers and Reagents/Sample Buffers &amp; Reagents ->MT::09e4dbf1-40c5-4a92-ace5-3ec31296ac7e##Life Science Research/Products/Protein Sample Preparation/Protein Sample Cleanup/ReadyPrep 2-D Cleanup Kit ->MT::bbf3a0be-188f-45c7-b81e-e9706ec77c02## Life Science Research/Solutions/Technologies/Protein Electrophoresis ->MTS::LUSOVO47B##Life Science Research/Solutions/Technologies/Western Blotting ->MTS::LUSPPAKG4##Life Science Research/Solutions/Technologies/2-D Electrophoresis ->MTS::LUSQG6LPT##Life Science Research/Solutions/Applications/Protein Separation and Analysis ->MTS::LUSOU3F7Q##Life Science Research/Solutions/Applications/2-D Electrophoresis and Analysis ->MTS::LUSQF04EH##Life Science Research/Solutions/Applications/Automated Electrophoresis and Analysis ->MTS::LUSQXB15##Life Science Research/Solutions/Technologies/Automated Electrophoresis System -Experion- ->MTS::LUSR04KG4## Life Science Research/Solutions/Technologies/2-D Electrophoresis/Protein Sample Preparation for 2-D Electrophoresis ->MTS::LUSQH6HYP## Eddie C Intro to Protein Sample Preparation 12/15/11 01:18 PM 12/15/21 01:19 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 en LSR /LSR/Technologies/Protein_Electrophoresis N 0 Cell Disruption /en-us/applications-technologies/applications-technologies/cell-disruption?ID=LUSP2D1FX Protein Solubilization /en-us/applications-technologies/applications-technologies/protein-solubilization?ID=LUSP3KFD4 Removal of Interfering Substances /en-us/applications-technologies/applications-technologies/removal-interfering-substances?ID=LUSP4LJSZ Sample Quantitation /en-us/applications-technologies/applications-technologies/sample-quantitation?ID=LUSP5JKAJ Introduction to Protein Electrophoresis /en-us/applications-technologies/applications-technologies/introduction-protein-sample-preparation?ID=LUSOVO47B

Protein sample preparation generally involves some or all of the following steps: cell disruption, extraction, removal of interfering compounds, changes of the physio/chemical properties and concentration of the sample. The quality of sample preparation can greatly affect the quality of the data that are generated. General guidelines and some of the most common methods for protein sample preparation are provided in this section. Protocols for sample preparation from mammalian tissue, plant leaves, microbial cultures, and for various buffer formulations are provided at the bottom of the page in the protocols tab.

Related Sections: Protein Electrophoresis Reagents Selection and Preparation

 

Protein Sample Preparation Workflow

Protein sample preparation workflow.

 

General Considerations

Due to the great diversity of protein sample types and sources, no single sample preparation method works with all proteins; for any sample, the optimum procedure must be determined empirically. However, the following general sample preparation guidelines should be kept in mind to avoid a number of pitfalls during sample preparation for protein electrophoresis (Posch et al. 2006).

  • Keep the sample preparation workflow as simple as possible (increasing the number of sample handling steps may increase variability)
  • With cell or tissue lysates, include protease inhibitors to minimize artifacts generated by proteolysis; protease inhibitors are generally not required for samples like serum or plasma
  • Solubilize proteins in a buffer that is compatible with the corresponding electrophoresis technique
  • Determine the amount of total protein in each sample using a protein assay that is compatible with chemicals in your samples
  • Use protein extracts immediately or aliquot them into appropriately sized batches and store them at –70°C to avoid freeze-thaw cycles
 

References

Posch A et al. (2006). Tools for sample preparation and prefractionation in two-dimensional gel electrophoresis. In Separation methods in Proteomics, Smejkal GB ed. (Boca Raton: CRC Press), pp 107–133.

 

Related Content

 
Literature
Number Description Download
3096 Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion Brochure, Rev B Click to download
6040 Electrophoresis Guide, Interactive PDF, Rev C Click to download
6194 Protein Sample Generation General Tips Click to download
6195 Human Cells Protocol Click to download
6196 Mammalian Tissue Protcol Click to download
6197 Plant Leaves Protocol Click to download
6198 Microbial Cultures Protocol Click to download
6199 Buffer Formulations Protocol Click to download
 
 
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The quality of sample preparation can greatly affect the quality of the data that are generated. General guidelines and some of the most common methods for protein sample preparation are provided in this section. Protocols for sample preparation from mammalian tissue, plant leaves, microbial cultures, and for various buffer formulations are provided at the bottom of the page in the <a href="#related_content">protocols</a> tab.</p> <p><strong>Related Sections</strong>: <a href="/evportal/destination/solutions?catID=LUSP8GD57">Protein Electrophoresis Reagents Selection and Preparation</a></p> Protein Sample Preparation Workflow <p><img class="stroke" src="http://www.bio-rad.com/webroot/web/images/1x1grey.gif" alt="" /></p> <p><img usemap="#Map" src="/webroot/web/images/lsr/solutions/technologies/protein_electrophoresis_blotting_and_imaging/protein_electrophoresis/technology_detail/pet13_img1.jpg" border="0" alt="" width="473" height="691" /> <map name="Map"> <area shape="rect" coords="5,4,475,143" href="/evportal/destination/solutions?catID=LUSP2D1FX"></area> <area shape="rect" coords="5,142,472,280" href="/evportal/destination/solutions?catID=LUSP3KFD4"></area> <area shape="rect" coords="4,278,475,442" href="/evportal/destination/solutions?catID=LUSP4LJSZ"></area> <area shape="rect" coords="4,441,476,580" href="/evportal/destination/solutions?catID=LUSP5JKAJ"></area> <area shape="rect" coords="5,579,474,692" href="#"></area> </map> </p> <p class="caption"><strong>Protein sample preparation workflow.</strong></p> <div class="top"><a href="#helptop">Back to Top</a></div> General Considerations <p>Due to the great diversity of protein sample types and sources, no single sample preparation method works with all proteins; for any sample, the optimum procedure must be determined empirically. However, the following general sample preparation guidelines should be kept in mind to avoid a number of pitfalls during sample preparation for protein electrophoresis (Posch et al. 2006).</p> <ul> <li>Keep the sample preparation workflow as simple as possible (increasing the number of sample handling steps may increase variability)</li> <li>With cell or tissue lysates, include protease inhibitors to minimize artifacts generated by proteolysis; protease inhibitors are generally not required for samples like serum or plasma</li> <li>Solubilize proteins in a buffer that is compatible with the corresponding electrophoresis technique</li> <li>Determine the amount of total protein in each sample using a protein assay that is compatible with chemicals in your samples</li> <li>Use protein extracts immediately or aliquot them into appropriately sized batches and store them at &ndash;70&deg;C to avoid freeze-thaw cycles</li> </ul> <div class="top"><a href="#helptop">Back to Top</a></div> References <p>Posch A et al. (2006). Tools for sample preparation and prefractionation in two-dimensional gel electrophoresis. In Separation methods in Proteomics, Smejkal GB ed. (Boca Raton: CRC Press), pp 107&ndash;133.</p> <div class="top"><a href="#helptop">Back to Top</a></div> <p><a name="related_content"></a></p> Protocols <table id="carttablealigned" class="literature_table" style="height: auto; width: 583px;" border="0" cellspacing="0" cellpadding="0"> <tbody> <tr> <td width="100">6194</td> <td width="350">Protein Sample Generation General Tips</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6194.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6195</td> <td width="350">Human Cells Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6195.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6196</td> <td width="350">Mammalian Tissue Protcol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6196.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6197</td> <td width="350">Plant Leaves Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6197.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6198</td> <td width="350">Microbial Cultures Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6198.pdf" target="_blank"><span>Click to download</span></a></td> </tr> <tr> <td width="100">6199</td> <td width="350">Buffer Formulations Protocol</td> <td class="pdf"><a class="pdf" href="/webroot/web/pdf/lsr/literature/Bulletin_6199.pdf" target="_blank"><span>Click to download</span></a></td> </tr> </tbody> </table> 3096 6040 /templatedata/internet/documentation/data/LSR/Literature/6040.xml Life Science Research/Products/Protein Sample Preparation/Protein Extraction/MicroRotofor Cell Lysis Kits ->MT::5e97c5be-3707-4c08-ae55-25e883f2ab99##Life Science Research/Products/Protein Sample Preparation/Protein Extraction/ReadyPrep Mini Grinders ->MT::0e0a3350-ceb8-4c52-9346-8f48ecee6e5a##Life Science Research/Products/Sample Quantitation/Protein Assay Kits and Cuvettes/<em>RC DC</em> Protein Assay ->MT::f58fdf1d-5d0b-4f31-93e2-f812bd7e7eab##Life Science Research/Products/Electrophoresis and Blotting/Protein Electrophoresis and Blotting/Electrophoresis Buffers and Reagents/Sample Buffers &amp; Reagents ->MT::09e4dbf1-40c5-4a92-ace5-3ec31296ac7e##Life Science Research/Products/Protein Sample Preparation/Protein Sample Cleanup/ReadyPrep 2-D Cleanup Kit ->MT::bbf3a0be-188f-45c7-b81e-e9706ec77c02## Life Science Research/Solutions/Technologies/Protein Electrophoresis ->MTS::LUSOVO47B##Life Science Research/Solutions/Technologies/Western Blotting ->MTS::LUSPPAKG4##Life Science Research/Solutions/Technologies/2-D Electrophoresis ->MTS::LUSQG6LPT##Life Science Research/Solutions/Applications/Protein Separation and Analysis ->MTS::LUSOU3F7Q##Life Science Research/Solutions/Applications/2-D Electrophoresis and Analysis ->MTS::LUSQF04EH##Life Science Research/Solutions/Applications/Automated Electrophoresis and Analysis ->MTS::LUSQXB15##Life Science Research/Solutions/Technologies/Automated Electrophoresis System -Experion- ->MTS::LUSR04KG4## Life Science Research/Solutions/Technologies/2-D Electrophoresis/Protein Sample Preparation for 2-D Electrophoresis ->MTS::LUSQH6HYP## Eddie C Intro to Protein Sample Preparation 12/15/11 01:18 PM 12/15/21 01:19 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 en LSR /LSR/Technologies/Protein_Electrophoresis N 0
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