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Protocols

All of the following protocols are in PDF format.

  • Rutgers
  • Antigen production  (Lab:Rutgers; Version:1; Added:August 1st at 9:14am)
    Express an antigen provided as a plasmid, biotinylate in vitro if desired, and purify the target protein.
  • JHU/CDI
  • Antigen coding gene induction and lysis  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Induction of expression of transfected gene and generation of a cell lysate for mAb validation
  • Cell line transfection  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Clone antigen coding gene into mammalian cell expression plasmid
  • ChIP-seq protocol  (Lab:JHU/CDI; Version:1; Added:January 27th at 12:00am)
    ChIP-seq protocol
  • Criteria for passing individual mAbs  (Lab:JHU/CDI; Version:1; Added:August 1st at 12:00pm)
    Criteria to determine whether an individual mAb has passed a primary validation (HuProt, OIRD, or Octet (r)) and a secondary validation (immunoprecipitation or immunoblotting/western blotting).
  • FL antigen  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Prepare full length antigen for immunization.
  • HeLa culture  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    HeLa cells culturing and plasmid transfection
  • HuProt validation  (Lab:JHU/CDI; Version:1; Added:August 9th at 2:51pm)
    Validate an antibody or other affinity reagent as monospecific using the HuProt human proteome array.
  • Hybridoma production  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Hybridoma production
  • IHC validation  (Lab:JHU/CDI; Version:1; Added:November 1st at 12:00am)
    Immunofluorescence Protocol
  • IP validation  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Validation of mAb by immunoprecipitation of target protein from cell lysate
  • Minichip hybridization  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Hybridize a binding reagent with a minichip.
  • Mouse immunization  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Immunize mice by intraperitoneal, intravenous, and footpad/hock routes.
  • Octet® Protocol  (Lab:JHU/CDI; Version:1; Added:July 31st at 9:23am)
    mAb affinity measurement using the Octet® system based on Bio-Layer Interferometry (BLI) technique
  • OIRD validation  (Lab:JHU/CDI; Version:1; Added:November 1st at 12:00am)
    OIRD Protocol
  • Protein purification  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Purify recombinant protein from 300 ml yeast culture.
  • Spiked IP Protocol  (Lab:JHU/CDI; Version:1; Added:March 5th at 2:06pm)
    Protocol for SpikeIP with Rutgers His‐tagged peptide/domains.
  • Western Blot Operations Procedures  (Lab:JHU/CDI; Version:1; Added:June 29th at 12:00am)
    Standard Operations Procedures For Western Blot
    Source: http://antibodies.cancer.gov/modules/site/pages/antibodies_detail_antigen_file.php?file_id=4&type=sop
    *Source link may direct you away from the Protein Capture website.
  • Yeast 300 ml culture  (Lab:JHU/CDI; Version:1; Added:September 30th at 4:01pm)
    Grow yeast in 300 ml culture for protein expression.