Question (25378) | Anti-KIF5A antibody (ab5628)

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Question

Dear Abcam technical team, I want to order proteins from your website, but there are some questions before order. (1) I want to experiment with conventional kinesin heavy chain, KIF5. But there are three different members of kinesin family 5. KIF5A, KIF5B and KIF5C. I know the function of each member, A and C involves in neuronal transport otherwise B involves in inner cell transport. But is there any structural difference between three members? If is, I have to choose specific member to experiment. (2) There are two types in KIF5A and three types in KIF5B. It seems that each has different domain, so I wonder there are functional or structural difference between types. Also, your company has two different types of KIF5C. One is just (938-957) domain and the other is His-tagged protein. His tagged kinesin has higher protein purity. Is there any reason to choose His tagged KIF5C except higher purity? (3) Is antibody that has specificity of my protein (e.g. KIF5A --> Anti-kinesin 5A antibody) attach to specific domain (like C-terminal) or it is just randomly attached? For my experiment, it is good for antibody to attach to C-terminal. In other words, is antibody has C-terminal specificity? (4) Do you sell biotinylated antibody for kinesin? I want antibody that has biotin coating but your website has no option or products for biotin coating. If you cannot understand my questions, please let me know which parts you cannot understand. Then I will send the more specific explanation for my questions. From

Answer

Thank you for contacting us and your interest in our products. 1.) Indeed, there is structural differences between those members. The sequences of the three members are not the same. I have made an alignment where you can see these differences. Please refer to the following link to see these differences: http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111011-085702-0894-35538860-oy To check out further parameters of these proteins, I can recommend to browse either the expasy site (http://web.expasy.org/ or http://web.expasy.org/protparam/) as well as one of the Bioinformatics institutes homepages, for example the EBI, http://www.ebi.ac.uk/. There are different Tools to predict the protein structures. 2.) We do have currently only one KIF5 protein in our catalogue, and this is the ab91695 Kinesin 5C protein (Tagged-His Tag) which is not a full length protein but a recombinant fragment corresponding to amino acids 500-957 of Human Kinesin 5C. Click here (or use the following: https://www.abcam.com/index.html?datasheet=91695). All the other products are indeed peptides and do correspond to the short amino acid sequence of the numbers in brackets. These peptides are too short to be able to have a function similar to the whole protein, however they can be used for blocking studies with the corresponding antibody. For example ab41784 is a peptide corresponding the sequence 938 to 957 of human KIF5C. Similarly for the other products: KIF5A: - peptide amino acid 1019-1032 is ab23133 and - peptide amino acid 1007-1027 is ab41781 KIF5B - peptide amino acid 376-396 is ab41782 - peptide amino acid 951-963 is ab23262 - peptide ab100817, a few amino acids derived from the sequence between amino acid 350 to 450 3.) Each antibody is different. I have therefore put together the following list: KIF5A, we do have two antibodies: -ab5628 is a polyclonal antibody directed against the C terminus of KIF5A, recognizing the sequences of amino acids 1007 to 1027 of KIF5A. This antibody has also been tested against the KIF5A protein of different species (human, mouse, rat and cow) as well as in immunocytochemistry, immunoprecipitation and Western blot. Click here (or use the following: https://www.abcam.com/index.html?datasheet=5628). -ab5958 which is a polyclonal antibody directed against the C terminal sequence (amino acids 1019 - 1032 of KIF5A). This antibody has however not been well characterized as yet, as you see it is one of our Fasttrack antibodies. Please let me know if you would like to know more about this. Click here (or use the following: https://www.abcam.com/index.html?datasheet=5958). KIF5B we have 7 antibodies currently: two of them are against a region in the C terminal: ab15705, corresponding to amino acids 951-963 Click here (or use the following: https://www.abcam.com/index.html?datasheet=15705). ab97762, corresponding to amino acids 901 - 963 Click here (or use the following: https://www.abcam.com/index.html?datasheet=97762). The other KIF5B antibodies are either N terminal (ab42492, immunogen amino acids 7-56), from the central region (ab5629, immunogen amino acids 376-396 and ab91033 region derived from amino acids 350 - 450) or are mouse monoclonal antibodies for which we do not know the epitope (ab110155 and ab11262) KIF5C we only have one antibody: ab5630 which has been raised against a C terminal sequence as well, the amino acids 938 - 957 of the human KIF5C. Click here (or use the following: https://www.abcam.com/index.html?datasheet=5630). 4.) Indeed we do currently not have one of these antibodies in biotinylated form. We do have however the Easylink conjugation kits, which are especially designed for easy conjugation of antibodies. We do have such kits also to perform biotin conjugation. You can find more information in this regards under the following website: www.abcam.com/easylink Again, please let me know if you wish more information about this. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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