Question (67389) | Anti-NMDAR2B antibody [NR2B] (ab28373)

Go to datasheet (ab28373)

Question

Dear Abcam manager


I just send you the mail again to ask about the 2nd antibody for fluoroscence WB detection.

I sent you the specification information on chemidoc system that we want to use last week but didn't get reply yet.

According to BioRad web sites, each color emission filter can detect below dyes.



RED: Cy5, Cy5.5, Alexa Fluor 647 / 680, DyLight 649 / 680, IRDye 680

GREEN: Cy3, Flamingo, Krypton, Pro-Q Diamond, Alexa Fluor 546, DyLight 549, Rhodamine

BLUE: Cy2, Coomassie Fluor Orange, Alexa Fluor 488, DyLight 488, Pro-! Emerald, Qdot 605 / 625 / 705

Answer

Thank you for your message and for your patience.
It is difficult to find three secondary antibodies in three colors that woudl be suitable, but I hope the following will help. Please check the excitation and emission wavelenghts and spectra to ensure they are suitable for the machine you are using. (I say this because some of the blue dyes they suggest go into the green range in the light spectra). I can recommend to review the online datasheets for further information.
Anti NMDAR1 antibody (ab77264, goat polyclonal)
Red fluorophore Secondary antibodies for detection of goat primary:
Donkey anti-Goat IgG H&L (PE) secondary antibody (ab7004)
Donkey F(ab')2 polyclonal Secondary Antibody to Goat IgG - H&L (PE), pre-adsorbed
https://www.abcam.com/index.html?datasheet=7004 https://www.abcam.com/index.html?datasheet=7004.
Anti-NMDAR2A antibody (ab14596, rabbit polyclonal)
Green fluorophore Secondary antibodies for detection of rabbit primary:
ab6798 Donkey anti-Rabbit IgG H&L (FITC) secondary antibody
https://www.abcam.com/index.html?datasheet=6798 https://www.abcam.com/index.html?datasheet=6798.

ab7079 Donkey anti-Rabbit IgG H&L (FITC) secondary antibody
https://www.abcam.com/index.html?datasheet=7079 https://www.abcam.com/index.html?datasheet=7079.
Anti-NMDAR2B antibody (ab28373, mouse monoclonal)
The following may fit into the blue channel on the instrument you are using, but I can suggest to please check this. The emission spectra is slightly into the green spectra.
ab96875 Donkey anti-Mouse IgG H&L (DyLight® 488) secondary antibody
https://www.abcam.com/index.html?datasheet=96875 https://www.abcam.com/index.html?datasheet=96875.
I am sorry we do not have any that are tested and covered by the 6 month guarantee in western blot to suggest on this occasion.
The other option is to directly conjugate the primary antibodies yourself with our Easylink kits. These are quick and easy to use. However, please note that we recommend the following for these kits:
The antibody to be conjugated should be in 10-50mM amine-free buffer within a pH range of 6.5-8.5. Modest concentrations of Tris buffer (up to 20mM) are also tolerated.
I can recommend to read the following important information and frequently asked questions about the kits before selecting a suitable antibody to conjugate:
https://www.abcam.com/index.html?pageconfig=resource&rid=13148
https://www.abcam.com/index.html?pageconfig=resource&rid=13156
I hope this will be helpful. Please do not hesitate to contact me if you have any further questions.

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