Parismita Kalita

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Questions related from Parismita Kalita

Any spectrophotometric method for determining selenocysteine content in protein?

Can anybody suggest a spectrophotometric method for detecting the presence of selenocysteine in a protein sample?

09 September 2016 3,369 1 View

What is the maximum concentration of imidazole that can be used to elute strongly bound protein from Ni-NTA column?

Lots of protein remains bound to the matrix even after elution with buffer containing 500mM Imidazole.

06 June 2016 1,331 3 View

Is it necessary to exclude stop codon in the primers sequence synthesized for clonning a gene into pET vectors with C-terminal His-tag?

I am using pET23(a) vector having a C-terminal His-tag to express a gene of 1.7kb. But as the reverse primer sequence includes stop codon , I think the his-tag expression is hindered. And...

03 March 2016 8,009 2 View

How to optimize his-tag protein binding to Ni-NTA column?

I have a His-tagged protein of ~70kDa, cloned into pET23(a) expression vector and the protein expresses in BL21 (DE3) cells when induced with 1mM IPTG and grown at 37C for 4hr after induction. The...

03 March 2016 315 9 View