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Questions related to Protein Purification Techniques
I have been trying to purify a GST-tagged eukaryotic protein from E. coli pLysS. I am getting it in low concentration along with a big blotch of non specific proteins.
03 March 2016 1,850 3 View
I am relatively new to western botting. When I first learned this procedure I used a 7.5% NEXT gel buffer blend, without the addition of a stacking gel. This is my first time running a...
03 March 2016 988 7 View
I expressed one protein in insect cell using bac-to-bac system. The protein was secreted in 293T cell. But it was stucked in cytoplasm. The protein was aggregated when it was lysised with...
02 February 2016 3,487 2 View
I have performed in-gel digestion protocol for Mass Spectrometry sample prepration and stored the trypsin digested gel and supernatant in -20C. Can the sample be stored for 1 week in such...
02 February 2016 491 4 View
Hi, I am working on recombinant protein production and I want to know whether I can remove urea and triton-X100 with dialysis. After the lysis of my cell pellet, I usually use 2M urea with...
02 February 2016 6,570 3 View
I am trying to run 2D gel. I am extracting protein from mammalian cell by using RIPA lysis buffer and then precipitate the protein with acetonitrile, then re suspend the protein in 250μl of...
02 February 2016 8,176 2 View
Hello, I am trying to find a good protocol for using these tubes. Does anyone know of any? I am targeting a DNA binding protein. I am new to this side of the sciences, so any help would be...
02 February 2016 9,603 2 View
The samples are Prawns muscle and are stored in ethanol 100% for a day at RT and later stored at -20ºC (2 weeks) I want to isolate RNA and DNA, just for a simple quantification and maybe later do...
02 February 2016 3,888 8 View
I am running PCR products of different size on an agarose gel, and I get bands that indicates products shorter than expected size. ( for your info agarose gel 1.5% and PCR product of expected size...
02 February 2016 3,995 2 View
what concentration of the buffer should I used inside the dialysis tube with protein and what concentration of the buffer should I used outside for the dialysis purpose? Can i use the same buffer...
02 February 2016 894 1 View
I am trying to isolate (unknown) proteins from a homogenized cell sample using phage (tomlinson library I+J) however the phage amounts we can create are too low to detect then isolate any protein...
02 February 2016 1,534 5 View
I am using TAMRA in a multiplex PCR( FAM/JOE/TAMRA/ROX/CY5 AB7500). The problem we have is that TAMRA seems to be suppressed by other 4 dyes severely. We are trying to find the reason. Should we...
02 February 2016 3,431 1 View
i needed to check cell signalling pathway, after treatment cells with some Conditioned media at some time curse . lysed and collected samples as usual. did bardfoard protein assay, samples had...
02 February 2016 3,856 6 View
We have been having consistently anomalous migration of high / higher MW DNA on our agarose gels. When running a linearized 5.3 kb plasmid, it's running above the 10 kb MW standard. However,...
02 February 2016 6,190 5 View
Hi, I have an unknown protein with the following info: Protein seq: M S D S E V N Q E A K P E V K P E V K P E T H I N L K V S D G S S E I F F K I K K T T P L R R L M E A F A K R Q G K E M D S L R...
02 February 2016 4,514 4 View
Hi I'm very new to this area and I'm just looking for where to start, I've been reading about ChIP and its variants. I am on the right lines, but the trouble I'm having is not knowing where to...
02 February 2016 8,197 3 View
How to isolate particular protein from seeds, can you tell me the simple protocol for that.
02 February 2016 4,526 2 View
Hello, I am trying to find interacting protein partners for a small nuclear intracellular domain (which is my protein of interest). My plan is to do IP with nuclear extract followed by Mass...
01 January 2016 5,249 3 View
Hi Everyone, I am going to purify a GST tagged protein from bacteria. I don't have a fixed angle rotor in the lab at the moment to centrifuge the samples after lysing the bacteria. I only have...
01 January 2016 3,954 5 View
I am doing pull down using His tagged protein A (28kDa) and an untagged protein B (47kDa). I incubate the proteins in presence and absence of a drug. In presence of drug protein A should...
01 January 2016 251 5 View
Dear Researchers, I performed Tyrosinase purification from bacteria broth medium , but I have difficulty to purify this enzymes because of the presence of melanin. Does someone have any idea on...
01 January 2016 8,049 2 View
What should be the optimum protein load (volume and concentration) when purifying a protein by sephadex G 100 , Column chromatography Any suggestions would be highly appreciated :)
01 January 2016 7,408 3 View
Does anyone have formulations for homogenization buffers for use with MAGPIX compatible: 1) protein multiplex panels 2) phosphoprotein multiplex panels. I'm interested in solubilizing...
01 January 2016 6,146 1 View
I need to remove the hemoglobin from the whole blood lysates and erythrocyte lysates for Mass spectrometry based proteomics. Since hemoglobin is most abundant protein which will mask and...
01 January 2016 6,805 5 View