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Technical training
 "Lecture and practice on cryopreservation of human iPS cells (vitrification method)"
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Inspection of antibody production

This inspection is for confirming the production of the following antibody isotypes from hybridoma cell lines.

  • Mouse monoclonal antibody class or subclass (IgG1, IgG2a, IgG2b, IgG3, IgM, IgA) and L chain(λchain, κ chain).
  • Rat monoclonal antibody class or subclass (IgG1, IgG2a, IgG2b, IgG2c, IgM) and L chain (κ chain).

1. Preparation
(1) Sample

  • Supernatant of mouse hybridoma cell lines
  • Supernatant of rat hybridoma cell lines

(2) Equipment

  • Microtubes

(3) Reagent

  • Mouse Monoclonal Antibody Isotyping Test Kit(Cat. No. MMT1)Bio-Rad
  • Rat Monoclonal Antibody Isotyping Test Kit(Cat. No. RMT1)Bio-Rad
  • PBS(-)

2. Method
(1) Take 50ul of sample to a microtube and add 450ul of PBS (-) for making 10-fold dilution, then stir it well.
(2) Pipette diluted 150ul sample into the development tube.
(3) After placing it for 30 sec at room temperature, vortex the tube softly and check if the reagent is dissolved.
(4) Insert the strip into the development tube with the red end at the bottom.
    Once both sides of blue positive control band become visible, remove the strip from the tube.

3. Interpretation of the result
Visually interpret the result of each class or subclass (and L chain).

Example: Mouse IgG2aκ