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Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 20 results for: RAP

Showing: 1 - 20 of 20 results

  1. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...Fisher 21028 Protein G GraviTrap columns, Cytiva 28-9852-55 or rProtein A GraviTrap columns, Cytiva 28-9852...and mix well. Uncap the Protein G Gravitrap or rProtein A Gravitrap columns and gently pour off the ethanol...Buffer Reservoirs to the top of the Gravitrap column. Pro-Tip Gravitrap columns have a finite binding capacity...the Gravitrap column to a clamp on a clamp stand. Use scissors to cut the bottom of the Gravitrap column...the Gravitrap columns at the bottom. Add 5 mL of Pierce IgG Elution Buffer to the capped Gravitrap column...430281 0.45 µm PES complete filtration unit, 500 mL, rapid-flow, VWR 73520-984 Syringes, 30 mL, VWR BD302832...upright at -20 °C Procedure Section 1: Affinity chromatography Equilibrate filtered tissue culture supernatant...

  2. CRISPR Library Amplification

    Type
    Protocol
    ...during the scraping process. Add each scrape into a 50 mL conical tube on ice pooling the scrapings of two...spreader and cold LB to scrape bioassay plates and remove bacteria. Use one scraper for all plates. Use two...mL cold LB to each plate for each scrape and use spreader to scrape plates. Pro-Tip Pushing motion is ... each tube on ice while scraping. Repeat addition of 10 mL cold LB and scrape for each plate until agar...spreading. Some spreaders have a sharp edge that can scrape plates more abrasively at a certain angle. Incubate...conical tubes on ice and ensure access to sterile scraper and cold LB. Pro-Tip Prepare one to two more conical...removing LB containing bacteria from plate). Pro-Tip Scraped bacteria in LB can clog the pipette either mix ...

  3. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... to collect the next fraction. The flow will be rapid at first and will progressively slow down. Option...methods improves infectious titer and yield." Gene therapy 6.6 (1999): 973-985 (Link opens in a new window... Rist Wolfgang, and Lamla Thorsten. Human Gene Therapy Methods. August 2015, 26(4): 147-157. doi:10.1089...

  4. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...microcentrifuge. Remove the gel from the plastic wrapper and rinse with deionized water. Gently remove the...Analyze . Select Gels . Select Plot Lanes . One graph per lane will appear with peaks representing each...

  5. Western Blot

    Type
    Protocol
    ...microtiter plate Chemiluminescence substrate Plastic wrap Primary antibody Secondary antibody Deionized water...BSA standard concentration versus absorbance. Extrapolate the total protein concentration of the sample...

  6. AAV Production in HEK293 Cells

    Type
    Protocol
    ...565-0010 (or for larger volumes 1000 mL 0.45 µm Rapid-Flow PES Filtration Unit, Nalgene 167-0045) Pro-...-Tip The pH of this solution will drift pretty rapidly upon addition of acid or base. Add only a few drops...

  10. Fluorescence Titering Assay

    Type
    Protocol
    ...associated with freeze-thaw. If using frozen virus, rapidly thaw the lentiviral aliquot on ice prior to use...

  11. General Transfection

    Type
    Protocol
    ...-Tip The pH of this solution will drift pretty rapidly upon addition of acid or base. Add only a few drops...

  12. Immunocytochemistry

    Type
    Protocol
    ...min at RT in the dark. Pro-Tip The plate can be wrapped in foil to block light. Remove the secondary antibody...

  13. Lentivirus Production

    Type
    Protocol
    ...first. Pro-Tip The pH of this solution will drift rapidly upon addition of acid or base. Add only a few drops...
Showing: 1 - 20 of 20 results