Direct conjugation for dual staining in immunohistochemistry

The simultaneous staining of two antigens can be a very powerful technique to study antigen expression but is traditionally time consuming and highly complex. This is due to a requirement for different primary antibody species and highly specific secondaries to eliminate cross-reactivity.

Lightning-Link® antibody labeling kits allow easy conjugation of primary antibodies with only 30 seconds hands-on time, eliminating the need for secondary antibodies and avoiding cross-species reactivity. 

The IHC data below shows that it is possible to obtain very good staining whilst avoiding the use of secondary antibodies, therefore saving time and reagent costs.


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Lightning-Link® kits were used  to conjugate mouse anti-human CD20 to HRP and rat anti human CD3 to Alkaline Phosphatase. Both antibodies were then used simultaneously to stain formalin fixed paraffin embedded sections of human lymph node. CD20 positive cells are visualised with DAB and are stained brown, and CD3 visualised with Fast Red and stained Red.


For more information on direct labeling for immunohistochemistry, take a look at the IHC application page and webinar. For any questions, please contact us at or locally at