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Showing: 1 - 13 of 13 results
  1. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...General Transfection Protocol Recombinant Antibody Purification Protocol Introduction Transfections allow...using Polyethylenimine Max as a transfection reagent. After transfection and expression, the recombinant...mL with >95% viability to proceed with transfection. Transfect the flask containing 108 mL cells: Transfer...feeding schedule: Thursday: Transfect cells. Friday (24 h post-transfection): Add 4% BCD Feed. Monday (...post-transfection): Add valproic acid to 3.75 mM, add 4% BCD Feed. Tuesday (120 h post-transfection): Add...Wednesday (144 h post-transfection): Add 4% BCD Feed. Thursday (168 h post-transfection): Harvest antibody...protocol for transfection of HEK293 cells to produce recombinant antibody. Protocols...
  2. General Transfection

    Type
    Protocol
    ...general method for transfecting mammalian cells using linear polyethylenimine. Transfections allow for transient... viral production) Day 1: Transfect Cells Day 2 (am): 18 h post transfection - Remove media, replace with...Carefully transfer the transfection mix to your HEK293T cells. Add the transfection mix dropwise, being ...protocol to transfect mammalian cells with your plasmid of interest. Protocols... Protocols General Transfection General...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral Service...adapted for different cell lines and different transfection reagents. Workflow Timeline Day 0: Seed HEK293T...
  3. Lentivirus Production

    Type
    Protocol
    ...lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol. This ...packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection. Remove media, replace...modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used...hydroxide 0.22 μm polyethersulfone (PES) filter Transfection grade DNAs Reagent Preparation DMEM Complete...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...5% CO 2 for ~20 h. Prepare a mixture of the 3 transfection plasmids into OptiPro SFM: Reagent Amount per...transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid DNA purification should ...
  4. AAV Production in HEK293 Cells

    Type
    Protocol
    ...cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment Class...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other transfection reagents may be used in this protocol, but their...their conditions must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing ... Pass and plate the cells the day before the transfection. Thaw a new vial of cells after 30 passages....to the incubator for 24 h–36 h. Proceed with transfection: Calculate the amount of each plasmid needed...Note that longer incubation times can reduce transfection efficiency. Add the OptiMEM + DNA + PEI solution...
  5. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Day 2: b. Perform the transfection in the late afternoon because the transfection mix should only be incubated...pLKO.1 can be introduced into cells via direct transfection, or can be converted into lentiviral particles...transient knockdown of protein expression, you may transfect plasmid DNA directly into the target cells. The...#12259 HEK-293T cells GenHunter: #Q401 FuGENE® 6 Transfection Reagent Roche Applied Biosciences: #11814443001...Producing Lentiviral Particles This protocol is for transfection in a 6 cm plate. The protocol can be scaled ... as needed. Day 1: a. For each plasmid to be transfected, plate 7×105 HEK-293T cells in 5 mL of media ...polystyrene tubes), make a cocktail for each transfection: 1 μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging...
  6. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...Virus Name Description Link to Video General Transfection Introduce plasmid DNA to mammalian cells Lentivirus...Produce lentivirus with a polyethyenimine (PEI) transfection protocol Fluorescence Titering Assay for Lentivirus...preparation Antibodies Name Description Link to Video Transfection Introduce plasmid DNA to mammalian cells to ...
  7. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... protein expression observed using transient transfection approaches, generating cell lines using lentiviral... stable cell line, as opposed to transiently-transfected cells, increases reproducibility, as it eliminates...variation associated with repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic...Different brands and FBS lots can promote or inhibit transfection. Test a variety of brands and FBS lot numbers...
  8. Fluorescence Titering Assay

    Type
    Protocol
    ...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...
  9. Immunocytochemistry

    Type
    Protocol
    ...you know expresses the protein, such as cells transfected with a plasmid to express the protein of interest...
  10. Colony Formation Titering Assay

    Type
    Protocol
    ...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...
  11. Western Blot

    Type
    Protocol
    ...you know expresses the protein, such as cells transfected with the protein of interest, and a negative ...
Showing: 1 - 13 of 13 results