ChromoBlog

The importance of blocking when using Nano-Secondaries for IF

Posted by Dr. Astrid Sitte on Jan 28, 2020 5:04:30 PM

Introduction

Blocking is an essential step during the preparation of a sample for immunofluorescence detection. Blocking improves the sensitivity by reducing nonspecific background and therefore increases image quality. Insufficient blocking results in higher background noise and over-blocking can even mask the specific signal.

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Topics: Nano-Secondaries, staining

What is one-step immunostaining?

Posted by Dr. Astrid Sitte on Jul 26, 2019 2:45:28 PM

Simultaneous immunostaining, also called one-step immunostaining vs. sequential immunostaining. Nano-Secondaries stain different primary antibodies equally well in one-step staining and sequential staining.

HeLa cells were immunostained with different primary antibodies and Nano-Secondaries Alexa Fluor® 647 (1:1,000, magenta). Cell nuclei were stained with DAPI (blue). Scale bar, 20 μm.

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Topics: live cell imaging, Secondary antibody, Nano-Secondaries, One-step immunostaining, staining

Important facts to know when using Nano-Secondaries

Posted by Klaus Herick on Jul 23, 2019 1:27:20 PM

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Topics: VHH, Nanobody, Secondary antibody, Nano-Secondaries, Alexa Fluor, One-step immunostaining

Advantages of recombinant Nano-Secondaries

Posted by Klaus Herick on May 23, 2019 4:10:12 PM

Nano-Secondaries are monoclonal Nanobodies that bind to primary antibodies in a species and subtype specific manner, i.e. Nano-Secondaries are very precise secondary antibodies. They are recombinantly produced in bacterial or other animal-free expression systems; in contrast, classical mono- or polyclonal secondary antibodies are traditionally produced using hybridoma technologies or are isolated from a host’s blood.

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Topics: Nanobody, Secondary antibody, Validation, recombinat expression, Nano-Secondaries

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