What is lowest amount of BSA protein that can be conjugated with citrate capped gold nanoparticles (20 nm) through direct binding (without using any linker)?I have prepared BSA stock solution of 0.1 mg/mL to 0.3 mg/mL (in water) and mixed them one by one at a ratio of 1:3 (BSA : AuNPs colloids) and found that the below 0.1 mg/mL conjugation, no shift in plasmon resonance wavelength (of AuNP) observed below 0.1 mg/mL?
I just want to know that is below 0.1 mg/mL conjugation and detection using spectrophotometer possible or not?
You do a saturation curve using set of AuNPs with increasing amount of BSA.
Surface plasmon increase can give you an idea about conjugation. But to be ceratin you should centrifuge the mixture and do HPLC with the supernatant. And do FTIR to confirm the attachment of BSA into it. So, for below 0.1 mg/mL you can use other techniques to be sure about conjugation.
Go through attached article you may find answer to your question. If any thing remains get back to me for further clarification.
Nanomolar concentration of BSA give better results.Nanomolar can't be prepared by directly weighing and adding Buffer to the BSA powder.It can be prepared with serial dilution.the Absorption spectra is comparable with nanoparticle concentration
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