A forum to address questions regarding methods of protein purification. | Contact experts in Protein Purification to get answers
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Questions related to Protein Purification
I am using IPTG induction to express my protein of interest. At the end of induction and cell lysis, I'll be re-suspending my pellet in 40ml buffer. My protein has a N-terminus Strep-II tag and...
06 June 2019 4,677 3 View
I have 5mM beta mercaptoethanol in my elution buffer used for protein purification. Will it interfere with an assay monitoring NADH to NAD+ oxidation spectrophotometrically? I am working with a...
06 June 2019 309 5 View
I need to perform the recombinant expression of a metalloenzyme. A 6xHis tag was planed to be fused at the C-terminus of the DNA for the protein purification. However, it might be very close to...
06 June 2019 6,512 3 View
Hi all, I am purifying a protein using anti-Flag magnetic beads. The protein purification is fine but I lose a lot of beads because they attach to the plastic (tips, tubes, falcon,..). Any...
06 June 2019 4,407 5 View
Dear researchers , I am simulating a cascode common source LNA (low noise amp) in 0.18um tech in cadence in ISM band of 902 -928 MHZ with the supply voltage of 0.9 v but , although all my...
31 May 2019 3,448 5 View
I am working with a protein-12kda; my vector contains 10X His tag My protocol for purification- NOTE- this is from 1 ml sonicated supernatant, used 100ul NiNTA beads. After I collect Flow...
24 May 2019 9,295 8 View
Can a human recombinant glycoprotein produced in mammalian cell culture lose part of its sialic acid residues due to changing conditions in the culture or the purification of the protein? Any...
18 May 2019 10,118 3 View
I express my protein in pTYB12 vector as fusion with intein and then try to purify on chitin beads. Typically, I load overnight at low flowrates (something like 0.3 ml/min). The loading buffer is...
09 May 2019 8,737 5 View
05 May 2019 8,558 9 View
I am looking for suggestions or comments from protein purification community, especially people who work with Con-A column chromatography. I am used Con-A Sepharose and Con-A agarose beads...
05 May 2019 10,016 6 View
I am trying to extract the protein content of extracellular bone matrix. After milling the frozen bone in nitrogen liquid, I have used RIPA buffer and HALT to extract the protein content, but...
05 May 2019 6,208 4 View
I am curious about nickel-bead cleaning. After native protein purification, I washed the nickel bead with 2 column volume of 2M Imidazole to remove the non-specific binding protein. But after...
05 May 2019 4,673 8 View
05 May 2019 5,317 3 View
05 May 2019 2,833 7 View
Cell-specific targeting via over expressed folate receptors or CD44 is a often reported in literature, but I am curious how this thing can be proved that actual binding happened. Literature often...
16 April 2019 8,335 2 View
I am doing some his-tagged protein purification and I would like to see if the chromatography works by running a standard. Is there a cheap histidine tagged protein people tend to use? Also the...
08 April 2019 9,480 4 View
Hi, I'm looking to further purify my protein by using cation exchange with Nuvia S resin. However, my lab doesn't have an IEX system. Is it possible to still carry out a cation exchange...
04 April 2019 7,077 5 View
I have some questions regarding the AKTA pure system, mainly about the chromatogram.
04 April 2019 2,848 4 View
04 April 2019 2,666 5 View
Hi, I am working with a protein that is typically secreted from the parasitic cells that I study. I am trying to express and purify it from HEK cells. Despite of using the mammalian signal...
02 April 2019 7,354 5 View
Can I use 450nm or any other wavelength instead of 340nm wavelength to measure mitochondrial ATP synthase activity (Complex V).
22 March 2019 4,712 3 View
I am treating neuroblastoma cells with AB42 protein (monomers and fibrils in separate experiments) and studying what happens after its uptake and concurrent interactions with intracellular...
19 March 2019 4,577 3 View
Hello everybody, I want to purify a 32 kDa protein with a C-terminal His-tag. I use the Histrap Crude FF and I elute with 500 mM Imidazol (no linear gradient) and I get a quite distinct peak with...
18 March 2019 7,867 2 View
Hi All, I have used MBP tag to help solubilize my target protein. It works really well. However, whenever I cleave off the His-MBP tag and try to clean up the sample by passing through another Ni...
09 March 2019 5,593 5 View