Re: direct labelling of primary antibodies **Vendor Reply**

[Kilgore, Jason A.]

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**Vendor reply**

Here at Molecular Probes we have a number of antibody kits that are more reliable.

For "no-purify" quick labeling kits, similar to the ones you linked to, we offer our Zip Alexa Fluor Kits.  Worth checking out.

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions
 

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Sylvie Le Guyader
Sent: Thursday, August 22, 2019 10:48 PM
To: [hidden email]
Subject: direct labelling of primary antibodies

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Dear list

Directly labelled antibodies are commonly used in FACS. The great advantages in using them are:

  *   No need to incubate with a secondary (expensive, time consuming, large, killing animals)
  *   No trouble finding secondaries raised in different species to avoid cross-binding when one labels with more than 3 antibodies.

Unfortunately for microscopy, one often wants to use an antibody that is not available with direct labelling. There are several companies that offer kits to directly label the primary antibody.

I would like to hear the experience of the list about direct labeling methods.

We have tried Flexistain<https://urldefense.proofpoint.com/v2/url?u=http-3A__kromnigon.com_flexistain_&d=DwIGaQ&c=q6k2DsTcEGCcCb_WtVSz6hhIl8hvYssy7sH8ZwfbbKU&r=MVp-2yJ1A-yQtCbZ-DE9xd0W6E7srQQpV-yioYjTLyQ&m=wBPWxP7x88k02KvAU_psro6OP0tD4lfcWcWVMffQ3R4&s=jg0t-H4PH79oQQGHpX0XCut9i9-8MmvIo0uA2O_W0_s&e= > and Mix-n-Stain<https://urldefense.proofpoint.com/v2/url?u=https-3A__biotium.com_product_mix-2Dn-2Dstain-2Dcf-2Ddye-2Dantibody-2Dlabeling-2Dkit_&d=DwIGaQ&c=q6k2DsTcEGCcCb_WtVSz6hhIl8hvYssy7sH8ZwfbbKU&r=MVp-2yJ1A-yQtCbZ-DE9xd0W6E7srQQpV-yioYjTLyQ&m=wBPWxP7x88k02KvAU_psro6OP0tD4lfcWcWVMffQ3R4&s=Szjpz3o16ruBs2ATAOSItY1j1PFVMNmpWwzOJm2Xu2c&e= >.

The first one requires Biotinylated primaries so that is a disadvantage if one already has an primary that works in the lab.

With Mix-n-stain we have had mixed results where the labeling worked nicely with some antibodies and not at all with others.

My questions are :

- what is your experience if any, with the kids mentioned above?

- could you recommend a way to directly label antibodies that works well for you?

Thanks

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD

Live Cell Imaging Facility Manager

Karolinska Institutet- Bionut Dpt

Hälsovägen 7C,

Room 7362 (lab)/7840 (office)

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