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.
Dear Tech Support Team,
Our customer is interested in ab154873 but he is concerned about high isoelectric point of his protein (PI=9). Please advise if ab154873 is suitable for the conjugation for this kind of the protein (it is a core of Hepatitis C).
Thanks in advance for your assistance and reply.
Asked on Mar 06 2013
Thank you for your enquiry regarding ab154873.
I can confirm that the high isoelectric point of the protein should not be a problem when using ab154873: EasyLink GOLD Conjugation Kit.
The main points to bear in mind about the protein buffer prior to labeling with this kit are:
● Protein must be purified
● Avoid amino acids (e.g. glycine)
● Avoid other primary amines (e.g. Tris)
● Avoid thiols (e.g. mercaptoethanol, DTT)
● Avoid carboxylic acids (e.g. EDTA)
Your customer should also note that the protocol will be the same as when labeling an antibody. However, if the protein is significantly larger or smaller than a typical IgG (about 160kD) they might want to explore varying ratios of protein.
The optimum amount of antibody or protein in this case (which will influence the number of antibody/protein molecules per particle) may be application-dependent and you may need to conjugate different amounts of antibody to optimize your assay.
I would advise your customer to download the protocol booklet (click on the blue hyperlink bottom section on the on-line datasheet) and read the document carefully.
The initial amount of antibody recommended corresponds to 10ug antibody per ml of 10 OD gold, which is about half of that normally used for passive (non-covalent) conjugations. However lower or higher concentrations can be explored as there is no risk of aggregation because of the protective surface coat.
I hope that this helps. If you need any further assistance in the future, please do not hesitate to contact me.
Answered on Mar 06 2013