Re: Well-liked 405 secondary antibodies ** commercial reply **

[Kilgore, Jason A.]

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** Commercial reply **

Hi, Joe,

I'm wondering how the results were disappointing. I'd be happy to troubleshoot offline.

But what I can add is that violet dyes in general tend to be dimmer and less photostable than many other wavelengths.  

In my testing, I have found the DyLight 405 is brighter and a bit more photostable than Alexa Fluor 405 when compared side-by-side, if that might help.

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions
[hidden email]
541-335-0353 - Tech Support
 

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Joe Lebowitz
Sent: Thursday, May 2, 2019 1:17 PM
To: [hidden email]
Subject: Well-liked 405 secondary antibodies

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Hello all,

We have had some reproducibly disappointing results with Alexa Fluor 405 tagged secondary antibodies.

Does anyone have a go-to secondary in this color other than the AF? We are trying to multiplex as many channels as possible and this is the only one we struggle to get good brightness/SNR with.

Thanks!

joe

--------------------------------

Joe Lebowitz
Ph.D. Candidate
NINDS Pre-doctoral Fellow
Khoshbouei Lab
UF Department of Neuroscience
[phone]

[Cammer, Michael]

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This is complicated by the 405 nm lasers on confocals. Alexa Fluor 350 works well on widefield fluorescent scopes with standard dapi blocks, but does not work at all on the confocal microscopes with 405 nm lasers.   Alexa Fluor 405 is  weak with our widefield scopes  that use a dapi block with ex at 365 nm (except for one with a 395 nm LED instead of a dapi block) whereas ideal with 405 nm laser using confocal.  We have to find out which scope people want to use before advising on a blue probe, but most people don't ask before they label.


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY  10016

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From: Confocal Microscopy List <[hidden email]> on behalf of Kilgore, Jason A. <[hidden email]>
Sent: Thursday, May 2, 2019 4:38:25 PM
To: [hidden email]
Subject: Re: Well-liked 405 secondary antibodies ** commercial reply **

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** Commercial reply **

Hi, Joe,

I'm wondering how the results were disappointing. I'd be happy to troubleshoot offline.

But what I can add is that violet dyes in general tend to be dimmer and less photostable than many other wavelengths.

In my testing, I have found the DyLight 405 is brighter and a bit more photostable than Alexa Fluor 405 when compared side-by-side, if that might help.

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions
[hidden email]
541-335-0353 - Tech Support


-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Joe Lebowitz
Sent: Thursday, May 2, 2019 1:17 PM
To: [hidden email]
Subject: Well-liked 405 secondary antibodies

CAUTION: This email originated from outside of the organization. Do not click links or open attachments unless you recognize the sender and know the content is safe.


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Hello all,

We have had some reproducibly disappointing results with Alexa Fluor 405 tagged secondary antibodies.

Does anyone have a go-to secondary in this color other than the AF? We are trying to multiplex as many channels as possible and this is the only one we struggle to get good brightness/SNR with.

Thanks!

joe

--------------------------------

Joe Lebowitz
Ph.D. Candidate
NINDS Pre-doctoral Fellow
Khoshbouei Lab
UF Department of Neuroscience
[phone]

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