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Now I am searching antibodies related to neurogenesis in the developing zebrafish. I am strongly interested in Neurogenin1, NeuroD4 and EphrinB3 antibodies in Abcam. As you know, it is hard to find good antibodies for zebrafish. Actually, a NeuroD4 anbody in Abcam may be useful for zebrafish due to homology of antigen. Could you give me feedback of possibility whether Neurogenin1, NeuroD4 and EphrinB3 antibodies have cross reactivity to zebrafish proteins? Zebrafish Neurogenin1: http://www.ensembl.org/Danio_rerio/Transcript/Sequence_Protein?db=core;g=ENSDARG00000056130;r=14:27712429-27713928;t=ENSDART00000078563 zebrafish NeuroD4: http://www.ensembl.org/Danio_rerio/Transcript/Sequence_Protein?db=core;g=ENSDARG00000003469;r=23:28586880-28589931;t=ENSDART00000145072 zebrafish EphrinB3b: http://www.ensembl.org/Danio_rerio/Transcript/Sequence_Protein?db=core;g=ENSDARG00000042277;r=7:22134347-22257954;t=ENSDART00000062003 Regards,
Asked on Oct 19 2011
Thank you very much for your inquiry. Indeed, it is difficult to find antibodies crossreactive with Zebrafish. I don't know whether I have already told you about our list of Zebrafish crossreactive antibodies? It can be found under the following link: www.abcam.com/zebrafish In regards to choosing the most probably crossreacting antibody for your three targets of interest, please find my comments here below. As for the previous antibody, please do not hesitate to ask for the discount code for testing these antibodies on Zebrafish (www.abcam.com/collaborationdiscount) I would like also to recommend, to chose antibodies which have been tested in the application or at least a similar application to which you will be using. Antibodies against Neurogenin 1: The ab89461 has a good alignment score and it seems likely that crossreactivity could happen. The score is 68% http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-113055-0569-24134208-pg https://www.abcam.com/index.html?datasheet=89461 Also the ab117847 has reasonable chances to detect the Zebrafish protein, as the full length human protein has been used as an immunogen and the score is 47%. http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-105747-0570-80392552-pg https://www.abcam.com/index.html?datasheet=117847 The two other antibodies, ab66498 and ab71818, are generated against peptides from N and C term, and as the homology there is not very good with the Zebrafish protein, I would recommend to test the other two antibodies first. NeuroD4 The immunogen of ab90484 has stretch conserved and I think it could work therefore. The score is 50% and the alignment can be found here: http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-110656-0883-28043163-pg https://www.abcam.com/index.html?datasheet=90484 The ab67168 has been produced with the full length human protein and the overall score is with 55% also relatively ok. http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-110901-0500-99792871-pg https://www.abcam.com/index.html?datasheet=67168 Ephrin B3 The overall protein alignment between the human and Zebrafish protein is relatively good, with a score of 54%.http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-111305-0838-36391612-pg The ab101699 has a reasonable good alignement and I could recommend to test this antibody therefore. The alignment is ********:::***: with a score of 73% http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111019-111510-0668-3570069-pg https://www.abcam.com/index.html?datasheet=101699 Of the other antibodies, I can also recommend to test either the ab28389 or the ab89474, as they have been produced with relatively long sequences and possibility of crossreactivity is therefore good as well. https://www.abcam.com/index.html?datasheet=28389 https://www.abcam.com/index.html?datasheet=89474 I would not recommend the ab53063 and for the ab103001 and ab106344 I do not know the immunogen sequences. If however these antibodies would interest you also in regards to their tested applications, I will be happy to ask the laboratory for prediction of the crossreaction. I hope this information is helpful. Please do not hesitate to contact me for the discount codes or for further information.
Answered on Oct 19 2011