{"product_id":"biofargo-cell-free-protein-synthesis-kit-max","title":"Biofargo Cell-Free Protein Synthesis Kit Max (Lyophilized)","description":"\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eDescription\u003c\/h2\u003e\n\u003cp\u003eBiofargo Cell-Free Protein Synthesis Kit Max produces protein in vitro from an \u003cem\u003eE. coli\u003c\/em\u003e lysate, with the redox conditions of the system already optimised for disulfide-bond formation. Transcription and translation are coupled in one tube, so a DNA or RNA template goes in and folded protein comes out without a living host.\u003c\/p\u003e\n\u003cp\u003eExpression starts within 1–2 hours and peak yield is reached in 8–24 hours, so a construct can be tested the same day it is built. Reported yields reach more than 3 mg\/mL. Because nothing has to stay alive, the reaction tolerates work that kills cells — toxic proteins, unnatural amino acids, labelled precursors and direct addition of cofactors.\u003c\/p\u003e\n\u003cp\u003eThe kit is supplied lyophilised, which keeps it stable at −20 °C for 12 months and lets it ship on blue ice rather than dry ice. Reactions run in tubes or 96-well plates and scale linearly from microlitres upward. A pJL1-sfGFP control plasmid is included, so a failed run can be told apart from a failed construct by eye. The Max formulation needs no separate disulfide-bond enhancer: the oxidising conditions are built into the system.\u003c\/p\u003e\n\u003cp\u003eFor Research Use Only. Not for use in diagnostic or therapeutic procedures, or in food or pharmaceuticals.\u003c\/p\u003e\n\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 90%;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eParameter\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eSpecification\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eProduct Name\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eBiofargo Cell-Free Protein Synthesis Kit Max (Lyophilized)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eCatalog Number\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eBF-P3002\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eExpression System\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\n\u003cem\u003eE. coli\u003c\/em\u003e cell-free system with optimised disulfide \/ redox conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003ePack Size\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e20 reactions (50 µL reaction scale)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eFormat\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eLyophilized\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eReported Yield\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eUp to \u0026gt;3 mg\/mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eTime to Protein\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eExpression within 1–2 h; peak yield at 8–24 h\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eTemplate\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003ePlasmid DNA, linear DNA, PCR product, RCA product or mRNA\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eTemplate Requirement\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eT7 promoter, ribosome binding site and T7 terminator; final concentration 5–10 µg\/mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eDisulfide-Bond Folding\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eOptimised within the system; no separate enhancer required\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eReaction Temperature\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e25–30 °C with vigorous shaking\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eReaction Volume\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eTypically 50–100 µL; scales linearly; tubes or 96-well plates\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003ePositive Control\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003epJL1-sfGFP control plasmid, 2 µg (Ex\/Em 485 \/ 528 nm)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eStorage\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 °C, valid 12 months. After reconstitution: −20 °C up to 1 month, −80 °C up to 12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eShipping\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eBlue ice\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eIntended Use\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eFor research use only\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eFeatures\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eProtein the same day\u003c\/strong\u003e — expression starts in 1–2 hours and peaks at 8–24 hours, against days for transformation, culture and induction.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eYields up to \u0026gt;3 mg\/mL\u003c\/strong\u003e, several times what a standard cell-free lysate delivers.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eDisulfide bonds handled in-system\u003c\/strong\u003e — the redox conditions are optimised in the formulation, so no separate disulfide-bond enhancer has to be bought or titrated.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eOpen reaction\u003c\/strong\u003e — nothing has to survive, so toxic proteins, unnatural amino acids, isotope labels and cofactors can be added directly.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eAny common template\u003c\/strong\u003e — plasmid, linear DNA, PCR product, RCA product or mRNA, with no cloning host required.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eLyophilized format\u003c\/strong\u003e — 12 months at −20 °C and shipping on blue ice rather than dry ice.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eScales and parallelises\u003c\/strong\u003e — reactions run linearly from microlitres up, in tubes or 96-well plates.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003esfGFP positive control included\u003c\/strong\u003e, so a failed reaction is distinguishable from a failed construct by eye.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eApplications\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eAntibody formats:\u003c\/strong\u003e Express full-length antibodies, nanobodies and antibody fragments that depend on correct disulfide pairing.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eCytokines and membrane proteins:\u003c\/strong\u003e Express disulfide-rich secreted and membrane proteins that fold poorly in a reducing cell-free system.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eConstruct screening:\u003c\/strong\u003e Test many designs in parallel in 96-well plates before committing to a cell-based expression run.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eToxic and difficult proteins:\u003c\/strong\u003e Express proteins that kill or are not tolerated by a living host.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eProtein engineering:\u003c\/strong\u003e Pair with Golden Gate or Gibson assembly and PCR or RCA templates for rapid design–build–test cycles.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eLabelled protein:\u003c\/strong\u003e Add unnatural amino acids or isotope labels directly to the reaction for structural and mechanistic work.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eEnzyme and assay reagents:\u003c\/strong\u003e Produce small amounts of active enzyme for assay development without a fermentation step.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eKit Components\u003c\/h2\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 90%;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eComponent\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eVolume\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eStorage\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eCell-free system solution A Max (Lyophilized)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e1 vial (300 µL after reconstitution)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 °C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eCell-free system solution B (Lyophilized)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e1 vial (600 µL after reconstitution)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 °C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eCFPS-Control Plasmid (pJL1-sfGFP)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e2 µg\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 °C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eStorage \u0026amp; Shipping\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cp\u003eShip on blue ice.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eStore the lyophilized kit at −20 °C. Valid for 12 months.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eAfter reconstitution, store at −20 °C for no more than 1 month, or at −80 °C for no more than 12 months.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eKeep reagents on ice while assembling reactions and avoid repeated freeze-thaw cycles.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eProtocol Overview\u003c\/h2\u003e\n\u003col\u003e\n\u003cli\u003e\u003cp\u003eReconstitute solution A with 300 µL and solution B with 500 µL of nuclease-free water, and mix thoroughly.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003ePrepare the template. Plasmids should be column-purified; PCR products need roughly 200 bp of flanking sequence upstream of the T7 promoter and downstream of the T7 terminator to protect the linear fragment from exonucleases. Quantify the template accurately before use.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eAssemble the reaction on ice in a nuclease-free tube. For a 50 µL reaction: 15 µL solution A (30%), 30 µL solution B (60%), template to a final 5–10 µg\/mL, and nuclease-free water to 50 µL.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eIncubate at 25–30 °C with vigorous shaking. Maximum yield is normally reached by about 8 hours; an overnight 16-hour reaction also works. Lowering the temperature to 25 °C helps some proteins fold, and the reaction time should then be extended.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eFor reactions above 100 µL, use a vessel with enough headspace or aeration — a shake flask at 200 rpm — since oxygen transfer limits yield at larger scale.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eTake 1 µL of total reaction for total protein, or of the supernatant for soluble protein, and run SDS-PAGE.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eRun the supplied pJL1-sfGFP plasmid alongside as a positive control. A successful reaction shows visible green fluorescence, and can be quantified on a plate reader at Ex\/Em 485 \/ 528 nm.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cstyle\u003e\n  .bf-rich .josh-faq { background: #fff; color: #000; font-family: \"IBM Plex Sans\", sans-serif; }\n  .bf-rich .josh-faq__inner { padding: 24px 0 40px; max-width: 100%; box-sizing: border-box; }\n  .bf-rich .josh-faq__item { border-bottom: 1px solid #e5e7eb; padding: 0; }\n  .bf-rich .josh-faq__item:first-child { border-top: 1px solid #e5e7eb; }\n  .bf-rich .josh-faq__question { width: 100%; margin: 0; padding: 20px 0; display: flex; align-items: center; justify-content: space-between; gap: 16px; color: #505052; background: none; border: 0; font: 700 16px\/1.4 \"IBM Plex Sans\", sans-serif; text-align: left; cursor: pointer; list-style: none; }\n  .bf-rich .josh-faq__question::-webkit-details-marker { display: none; }\n  .bf-rich .josh-faq__question:hover { color: #505052; }\n  .bf-rich .josh-faq__icon { position: relative; width: 20px; height: 20px; flex: 0 0 20px; transition: transform .25s ease; }\n  .bf-rich .josh-faq__icon::before, .bf-rich .josh-faq__icon::after { position: absolute; content: \"\"; background: currentColor; border-radius: 2px; transition: opacity .25s ease, transform .25s ease; }\n  .bf-rich .josh-faq__icon::before { top: 50%; left: 0; width: 100%; height: 2px; transform: translateY(-50%); }\n  .bf-rich .josh-faq__icon::after { top: 0; left: 50%; width: 2px; height: 100%; transform: translateX(-50%); }\n  .bf-rich .josh-faq__item[open] .josh-faq__icon::after { opacity: 0; transform: translateX(-50%) rotate(90deg); }\n  .bf-rich .josh-faq__answer { max-width: 860px; padding: 0 0 20px; color: #505052; font: 400 16px\/1.6 \"IBM Plex Sans\", sans-serif; }\n  .bf-rich .josh-faq__answer p { margin: 0 0 15px; color: #505052; font: inherit; }\n  .bf-rich .josh-faq__answer p:last-child { margin-bottom: 0; }\n  .bf-rich .josh-faq__answer ul { margin: 8px 0 12px; padding-left: 22px; }\n  .bf-rich .josh-faq__answer li { margin-bottom: 6px; font: inherit; list-style-type: disc; }\n  @media (max-width: 767px) { .bf-rich .josh-faq__inner { padding: 24px 0 40px; } .bf-rich .josh-faq__question { padding: 16px 0; font-size: 15px; } .bf-rich .josh-faq__answer { font-size: 15px; } }\n\u003c\/style\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eFrequently Asked Questions (FAQs)\u003c\/h2\u003e\n\u003cp\u003eCommon questions about Biofargo Cell-Free Protein Synthesis Kit Max (Lyophilized).\u003c\/p\u003e\n\u003cdiv class=\"josh-faq\"\u003e\u003csection class=\"josh-faq__inner\"\u003e\u003cdiv class=\"josh-faq__list\"\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow long does a reaction take?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eExpression begins within 1–2 hours. Maximum yield is normally reached by about 8 hours at 25–30 °C, and an overnight 16-hour reaction also works. Running at 25 °C helps some proteins fold but needs a longer incubation.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhat yield should I expect?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eThe kit is specified at up to more than 3 mg\/mL. Actual yield varies a great deal with the target protein, so treat the figure as the system's capability rather than a guarantee for a given construct. Run the supplied sfGFP control to confirm the system is performing before judging a difficult target.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhat templates can I use?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003ePlasmid DNA, linear DNA, PCR products, RCA products from phi29 amplification, or mRNA. The template needs a T7 promoter, a ribosome binding site and a T7 terminator, at a final concentration of 5–10 µg\/mL.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhich vectors work, and which do not?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eThe included pJL1-sfGFP can be used as a cloning vector, and pET-series plasmids that carry the T7 promoter without the lactose operon — such as pET-9a and pET-23a — are compatible. Plasmids containing the lac operon, pET28a among them, significantly reduce yield and are not recommended.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow should I prepare a PCR-product template?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eDesign the forward primer about 200 bp upstream of the T7 promoter and the reverse primer about 200 bp downstream of the T7 terminator. Those flanks protect the linear fragment from endogenous exonucleases, and the product can then be used without purification.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eDoes plasmid purity matter?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eYes, more than for most applications. Use a high-purity plasmid prep with a separate deproteinization wash buffer and no RNase A carryover, and elute in nuclease-free water. Plasmid that has not been column-purified cannot be used directly in the reaction.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eCan it make disulfide-bonded proteins?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eYes. The Max formulation has the oxidising and redox conditions optimised within the system, so full-length antibodies, nanobodies, antibody fragments, cytokines and membrane proteins can be expressed without adding a separate disulfide-bond enhancer.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow do I scale the reaction up?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eReactions scale linearly. A typical run is 100 µL in a 2 mL microcentrifuge tube. Above that, oxygen transfer starts to limit yield, so use a vessel with sufficient headspace or aeration — a shake flask at 200 rpm.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow do I know the reaction worked?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eRun the supplied pJL1-sfGFP control plasmid. A successful reaction fluoresces green and can be quantified on a plate reader at Ex\/Em 485 \/ 528 nm. For the target protein, run 1 µL of total reaction or supernatant on SDS-PAGE.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow is it stored and shipped?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eIt ships on blue ice and is stored lyophilized at −20 °C, where it is valid for 12 months. Once reconstituted, keep it at −20 °C for up to 1 month or at −80 °C for up to 12 months.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eCan it be used for clinical or diagnostic purposes?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eNo. This product is for research use only and is not intended for human, diagnostic or therapeutic use.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003c\/div\u003e\u003c\/section\u003e\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cscript type=\"application\/ld+json\"\u003e{\"@context\": \"https:\/\/schema.org\", \"@type\": \"FAQPage\", \"mainEntity\": [{\"@type\": \"Question\", \"name\": \"How long does a reaction take?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Expression begins within 1\u0026ndash;2 hours. Maximum yield is normally reached by about 8 hours at 25\u0026ndash;30 \u0026#176;C, and an overnight 16-hour reaction also works. Running at 25 \u0026#176;C helps some proteins fold but needs a longer incubation.\"}}, {\"@type\": \"Question\", \"name\": \"What yield should I expect?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"The kit is specified at up to more than 3 mg\/mL. Actual yield varies a great deal with the target protein, so treat the figure as the system's capability rather than a guarantee for a given construct. Run the supplied sfGFP control to confirm the system is performing before judging a difficult target.\"}}, {\"@type\": \"Question\", \"name\": \"What templates can I use?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Plasmid DNA, linear DNA, PCR products, RCA products from phi29 amplification, or mRNA. The template needs a T7 promoter, a ribosome binding site and a T7 terminator, at a final concentration of 5\u0026ndash;10 \u0026#181;g\/mL.\"}}, {\"@type\": \"Question\", \"name\": \"Which vectors work, and which do not?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"The included pJL1-sfGFP can be used as a cloning vector, and pET-series plasmids that carry the T7 promoter without the lactose operon \u0026mdash; such as pET-9a and pET-23a \u0026mdash; are compatible. Plasmids containing the lac operon, pET28a among them, significantly reduce yield and are not recommended.\"}}, {\"@type\": \"Question\", \"name\": \"How should I prepare a PCR-product template?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Design the forward primer about 200 bp upstream of the T7 promoter and the reverse primer about 200 bp downstream of the T7 terminator. Those flanks protect the linear fragment from endogenous exonucleases, and the product can then be used without purification.\"}}, {\"@type\": \"Question\", \"name\": \"Does plasmid purity matter?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Yes, more than for most applications. Use a high-purity plasmid prep with a separate deproteinization wash buffer and no RNase A carryover, and elute in nuclease-free water. Plasmid that has not been column-purified cannot be used directly in the reaction.\"}}, {\"@type\": \"Question\", \"name\": \"Can it make disulfide-bonded proteins?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Yes. The Max formulation has the oxidising and redox conditions optimised within the system, so full-length antibodies, nanobodies, antibody fragments, cytokines and membrane proteins can be expressed without adding a separate disulfide-bond enhancer.\"}}, {\"@type\": \"Question\", \"name\": \"How do I scale the reaction up?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Reactions scale linearly. A typical run is 100 \u0026#181;L in a 2 mL microcentrifuge tube. Above that, oxygen transfer starts to limit yield, so use a vessel with sufficient headspace or aeration \u0026mdash; a shake flask at 200 rpm.\"}}, {\"@type\": \"Question\", \"name\": \"How do I know the reaction worked?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Run the supplied pJL1-sfGFP control plasmid. A successful reaction fluoresces green and can be quantified on a plate reader at Ex\/Em 485 \/ 528 nm. For the target protein, run 1 \u0026#181;L of total reaction or supernatant on SDS-PAGE.\"}}, {\"@type\": \"Question\", \"name\": \"How is it stored and shipped?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"It ships on blue ice and is stored lyophilized at \u0026minus;20 \u0026#176;C, where it is valid for 12 months. Once reconstituted, keep it at \u0026minus;20 \u0026#176;C for up to 1 month or at \u0026minus;80 \u0026#176;C for up to 12 months.\"}}, {\"@type\": \"Question\", \"name\": \"Can it be used for clinical or diagnostic purposes?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"No. This product is for research use only and is not intended for human, diagnostic or therapeutic use.\"}}]}\u003c\/script\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eRelated Products\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\u003cp\u003e\u003ca href=\"\/products\/biofargo-cell-free-protein-synthesis-kit\"\u003e\u003cstrong\u003eBiofargo Cell-Free Protein Synthesis Kit\u003c\/strong\u003e\u003c\/a\u003e — the standard formulation, for proteins that do not depend on disulfide bonds.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cp\u003e\u003ca target=\"_blank\" rel=\"noopener\" href=\"https:\/\/biofargo.com\/pages\/contact\"\u003e\u003cstrong\u003eRequest Datasheet\u003c\/strong\u003e\u003c\/a\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n","brand":"Biofargo","offers":[{"title":"Default Title","offer_id":53432384618677,"sku":"BF-P3002","price":240.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/biofargo.com\/products\/biofargo-cell-free-protein-synthesis-kit-max","provider":"Biofargo","version":"1.0","type":"link"}