Human peripheral blood mononuclear cells stained with ab235128 (right) or mouse IgG1κ (ab170190) isotype (left). Human peripheral blood mononuclear cells were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab235128) or mouse IgG1κ (ab170190) (1x10⁶ in 100 µl at 1 µg/ml) for 30 min on ice.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (ab150177) was used at 1/2000 dilution for 30 min at 4°C. The cells were simultaneously stained with CD19 antibody.

Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on viable lymphocytes.

IHC image of human Kappa chain staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BOND

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.