Bio-rad Mouse Typer Isotyping Kit Manual do Utilizador descarregar pdf (Página 12)

Problem Probable Cause Recommended Solution

C. Monoclonal 1.Impure sample. 1. Use fresh source of cul-

exhibits multiple ture supernatant. Purify

sub-isotype (i.e. media or ascites fluid.

5-7

assay shows one 2.Sample too con- 2. Dilute supernanants and

clone to be both centrated ascites fluid, repeat test.

an IgG1 and 3.Sample is polyclonal. 3. Re-clone hybridoma cells

IgM). by limited dilution,

and

repeat assay.

Notes to Troubleshooting Guide

1. Activity Test for Horseradish Peroxidase Substrate Solution.

Combine 1.0 ml of substrate solution with 10 ml of antibody

conjugate. If no color develops in 5 minutes, substrate solution

is at fault.

2. Activity Test for Antibody Conjugate Solution.

Combine 1.0 ml of horseradish peroxidase substrate solution

(tested above) and 1.0 ml of diluted antibody conjugate

solution. If no color develops in 5 minutes, the conjugate is

suspect. Repeat procedure using fresh conjugate antibody

dilution.

3. Activity Test for Monoclonal Antibody Solution.

Use RID, Ouchterlony precipitation, or ELISA assay to

determine reactivity. Repeat procedure with more

concentrated monoclonal antibody solution.

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Bio-rad Mouse Typer Isotyping Kit Manual do Utilizador Página 12