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Low protein signal during immobilisation
- biber
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10 years 4 months ago #1
by biber
Low protein signal during immobilisation was created by biber
Hİ everybody,
I use Bionavi210A SPR. I'm still learning. I try to immobilze HSA with amino coupling on CMD (=CM5) chip. But my protein signal is very low. My coupling buffer is 10 mM Na Acetate pH: 5.2. HSA is globulin and fatty acid free. I tried a lot of different chemical such as different protein (BSA), different EDC and NHS, Different buffer ( 5 mM MES pH: 5.0) but nothing changes. I also tried different concentraion of proteins. I sent you my sensogram results.
I sent you my sensogram results for BSA and HSA .
Do you have any recommadation to solve this problem?
I use Bionavi210A SPR. I'm still learning. I try to immobilze HSA with amino coupling on CMD (=CM5) chip. But my protein signal is very low. My coupling buffer is 10 mM Na Acetate pH: 5.2. HSA is globulin and fatty acid free. I tried a lot of different chemical such as different protein (BSA), different EDC and NHS, Different buffer ( 5 mM MES pH: 5.0) but nothing changes. I also tried different concentraion of proteins. I sent you my sensogram results.
I sent you my sensogram results for BSA and HSA .
Do you have any recommadation to solve this problem?
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- MQAdmin03
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10 years 4 months ago #2
by MQAdmin03
Replied by MQAdmin03 on topic Low protein signal during immobilisation
Hi Biber,
Not sure what i see in your sensorgrams (immobilization or experiment; please explain) but i have some suggestions.
- take a fresh equilibrated sensor chip
- inject the HSA solution in different pH buffers ( www.sprpages.nl/immobilization/ligand-pre-concentration.html )
- take the pH solution with the "best" pre concentration properties.
- then make fresh NHS and EDC according to the Biacore protocol or by the provided BioNavis protocol.
- Try again.
Not sure what i see in your sensorgrams (immobilization or experiment; please explain) but i have some suggestions.
- take a fresh equilibrated sensor chip
- inject the HSA solution in different pH buffers ( www.sprpages.nl/immobilization/ligand-pre-concentration.html )
- take the pH solution with the "best" pre concentration properties.
- then make fresh NHS and EDC according to the Biacore protocol or by the provided BioNavis protocol.
- Try again.
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