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  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...and diluted cell culture supernatant into the Gravitrap column. Collect the flow through in a 250 mL bottle...

  2. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ... 1mm thick, 10-well, Invitrogen NP0321BOX 20X MOPS SDS running buffer, Invitrogen NP0001 4X NuPage sample...buffer, Invitrogen NP0007 10X NuPage sample reducing agent (500 mM dithiothreitol, DTT), Invitrogen NP0009...NP0009 PageRuler Plus Prestained protein ladder, Invitrogen 26619 Pipette tips, 10 µL, VWR 76322-132 Pipette...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...serum-free media Invitrogen: #31985 Dulbecco’s Modified Eagle Medium (DMEM) Invitrogen: #11995 Fetal Bovine...Bovine Serum (FBS) Invitrogen: #16000 Penicillin/Streptomycin Invitrogen: #15140-122 Polypropylene tubes... buffer NEB #B0202S DH5 alpha competent cells Invitrogen #18258-012 Qiaquick gel extraction kit Qiagen...homologous recombination of unstable regions, such as Invitrogen Stbl3™ or NEB Stable cells. This will ensure ...

  4. Ligation Independent Cloning

    Type
    Protocol
    ...exonuclease function of the enzyme and shift its activity back to polymerase. In our case, dGTP will be ...protocol), causing the enzyme to perform exonuclease activity until the first "G" in the sequence. Because the...proceeding, as they will interfere with the exonuclease activity of T4 Pol in the following step. This may be accomplished...

  5. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ... However, keep in mind that restriction enzyme activity is determined under ideal conditions with very...their recognition sites. This is due to "Star Activity" and can happen for a variety of reasons, including...opens in a new window) NEB's website about star activity . If you are digesting a large number of plasmids...

  6. Gibson Assembly Protocol

    Type
    Protocol
    ...joined in a single isothermal reaction. With the activities of three different enzymes, the product of a ...ssDNA ends from the ssDNA-specific endonuclease activity of T5 Exonuclease and furthermore reduces secondary...
Showing: 1 - 9 of 9 results