For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
If you continue without changing your cookie settings, we'll assume you’re happy with this.
I have the following inquiries about the EasyLink Conjugation Kit: 1. Will the unbounded fluorophore mix or free labels (i.e. EL-FITC mix added to antibody) cross-reacts with sample added? 2. How long can the conjugated antibody be stored and at what temperature? Some antibodies require storage -20 deg Celcius, what is the storage temperature if conjugated with EasyLink labels? 3. My customer intends to use anti-STRO1 (ab102969) which is of isotype IgM with EasyLink FITC Conjugation kit. Would the labeling kit compatible for use with antibodies from different isotypes i.e. IgM, IgG2a, IgG2b, etc.?
Asked on Jan 05 2012
Thank you for your enquiry. Here are the answers to your questions. 1) The Easylink kits are designed to give low level of free label at the end of the reaction. Therefore, there will be no or very low unbounded fluorophore when the conjugated antibody is used on the sample. However, I would strongly recommend performing proper washing after antibody incubation (at least 5-10 min for 3 times), which will also help eliminate any non-specific binding and high background in the experiment. 2) There are no guidelines for storage after conjugation with the kit. In general, the conjugated antibody can be stored at 4°C for up to 6 months. Please include antimicrobial agents and/or stabilizers (e.g. azide, BSA, glycerol, etc.). Some antibodies can be stored at -20°C for longer storage over a year. However, please bear in mind that the storage temperature and duration is not dependent on the conjugation but on the antibody being used. Therefore, if the primary antibody can be stored at 4°C or -20°C, then the conjugated version can also be stored at that temperature. Please refer to the primary antibody storage instructions. 3) The conjugation kits will work with any of the antibody isotypes (IgM, IgG, IgA, IgD, IgE, IgG2a, IgG2b, etc). The labeling chemistry involves free amines present in lysines and at the N-terminus of the antibody. All antibodies have multiple free amine groups, therefore it will work even with ab102969 (IgM isotype). However, one thing that is not compatible is that ab102969 is of ascites purity, which is not recommended for use with Easylink kits. There is a need to purify the antibody using a purification kit like ab109208 Antibody Serum Purification kit (1 purification) before using an Easylink kit. I hope this is the information you are looking for. If everything is what your customer expect, then just simply choose the desired conjugation from the list of Easylink kits (https://www.abcam.com/index.html?pageconfig=resource&rid=13148). If there is anything else that I can help, please let me know.
Answered on Jan 05 2012