Hello everyone,
I am looking for antibody against EYFP. I have searched on Sigma, Thermoscientific, Abcam, and Cell signaling technology for EYFP antibodies. However, the search results show antibodies for GFP. I have the following doubts here:
1. Do anti-GFP antibody works for YFP as well?
2. Are antibodies for YFP and EYFP are different?
Anyone who has used the anti-YFP antibody, please suggest a reliable source/company for the antibody purchase.
Looking forward to hearing from you!
Thanks in advance
Shubham
Shubham,
Polyclonals raised against the whole GFP probably work, as these fluorescent proteins have much of the peptide sequence in common (AFAIR). For a monoclonals or peptide antibodies, you need to check the peptide sequence they recognize.
If in doubt or no reliable information is available, try a polyclonal.
GFP and YFP even though they share the spectrum for a certain range do not have the same polypeptide sequence. So as per my understanding anti GFP, might not entirely work for YFP.
EYFP and YFP should still be the same since EYFP should mean the sequence of YFP is present in more than one repeat (enhanced YFP). There are several companies that sell anti YFP. (For example- https://www.lsbio.com/antibodies/eyfp-antibody-clone-mix-wb-western-ls-c387458/399559). You could try those.
Hi Shubham,
I agree with the others.
The aa sequences of GFP and EYFP/YFP are very similar.
But this nevertheless depends on the individual antibody and the peptide the antibody was raised against. But I know at least two different polyclonal anti-GFP antibodies that work well against EYFP (can give you of course details - can look it up next week).
If you already have one in mind you may want to use, I would check whether the selling company provides the peptide sequence the antibody was raised against and check whether this overlaps with YFP/EYFP.
Thank you for your valuable suggestions Urs Mörbe Debadrita Pal and Wolfgang Schechinger. Unfortunately, I am unable to decide which one to go for. I missed adding that we are looking for antibody application in Immunofluorescence. Urs Mörbe It will be great if you can share the details about the polyclonal anti-GFP antibodies which works against EYFP.
Thanks
Shubham
Shubham Dwivedi ,
To echo Wolfgang Schechinger , here is one example from a company below: https://www.abcam.com/gfp-antibody-ab6556.html Read on....
- Product: GFP antibody (ab6556)
- Rabbit *polyclonal* to GFP
- Specificity of product: GFP antibody (ab6556) is reactive against all variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP, YFP, CFP, RFP and EGFP.
Working well for me are:
Invitrogen anti-GFP (polyclonal chicken) cat no. A10262
--> Would recommend an anti chicken - AF488 secondary
Takara living colors anti-GFP (polyclonal rabbit) car no. 632592
--> Would recommend an anti rabbit - AF488 secondary
The rabbit one has maybe a little more background in my hands, but I would buy in depending on what secondaries you´d have available anyway already.
Hello everyone,
Just to update you that I have used anti-GFP (Cat no #55494S) from Cell Signaling Technology, Host species: Mouse, Reactivity: all.
The secondary antibody used: Alexa 568 antimouse (Cat no #A11004) from Invitrogen. The antibody is able to detect the eYFP labeling in the Nerve, DRG, and Spinal cord as tested in our experiment.
Thank you for all the help provided here Urs Mörbe Yuan-Yeu Yau Debadrita Pal Wolfgang Schechinger.
Regards
Shubham
Shubham Dwivedi Adding another antibody which has worked beautifully in our hands to detect YFP across multiple species. Its the Anti-Green Fluorescent Protein Antibody from Aves Lab. SKU: GFP-1020.
Being an IgY harvested from chickens it produces very little background. Works at 1:1000 diluted in 3% BSA in 0.1% PBSTx for floating sections tested upto 100um thick and overnight incubation at 4 degrees. We do standard secondary amplification with A488 anti-chicken antibody for 2h at RT using the same diluent and 3 washes in between with 0.1% PBSTx. Hasn't failed yet. Best of Luck!
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA