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Questions related to Biochemical Techniques
I have two proteins which have almost the same M.Wt. They also have isoelectric points (7.1 and 7.75). Can I separate them with Ion exchange chromatography(IEC)? What is the PI range by which Ion...
10 October 2014 10,021 7 View
I am using Clustal Omega to align my protein sequences. That works really well, and I get residues that are conserved. But it does not calculate the percentage of identical residues. I tried to...
10 October 2014 3,657 2 View
We are attempting to culture various cell time at narrow ph margins. Ph of media is optimized to ranges of -+ 0.05. This can be attained at a constant level for several weeks at a time however we...
10 October 2014 733 2 View
I have tried to dissolve L-Cysteine in distilled water but I found it insoluble in distilled water. While googling I found that by mild heating and stirring or by dissolving L-cysteine in a water...
10 October 2014 6,990 10 View
I'm conducting a experiment where I need to detect cytokines and measure concentration of cytokines in supernatants collected from cell culture. Does anyone have experience on this issue? I...
10 October 2014 8,655 26 View
My group have nicely established glutardehyde cross-link to study the complex formation of the protein of our interest.We are working on both total tissue/cell extracts or recombinant proteins,...
10 October 2014 2,670 4 View
When designing the sequenom primer, the question is like this . Can anyone explain about it?
10 October 2014 4,150 1 View
I am trying to get only whole genome DNA from some old samples. When I run a gel I'm getting my band of genomic DNA but also a large amount of smearing lower on the gel likely due to sample...
10 October 2014 7,982 5 View
Hello !Does anyone know a good cleaning protocol to re-use C18 SPE material ?. We are currently using a glass column and want to recycle C18 solid phase, but we are not sure which procedure we...
10 October 2014 6,690 11 View
I am about to start some new experiments that rely on TAT-Cre (from Millipore) to delete a gene of interest in cell culture in order to study the impact on cell differentiation and gene...
10 October 2014 6,517 6 View
I try to do qPCR for dark-operative protochlorophyllide reductase (DPOR) subunits (chlL, chlB, chlN) in Norway spruce (Picea abies Karst.). I have the specific primers for these subunits and also...
10 October 2014 624 6 View
I am looking for a protein secreted by BMDCs over 24h. I need allot of cells (2 * 10^6) so i also need allot of medium (i am using RPMI + 10% FBS (R10)). However, turns out my protein of interest...
10 October 2014 6,387 2 View
Can anyone please tell me the effective concentration of trypsin used for trypsinization of H9c2 cells? I had used 1x trypsin from Sigma and got good results. But now I got 10x trypsin and it did...
10 October 2014 5,440 3 View
We have both paraffin- waxed tissue samples and blood from patients We would like to know what is the best method to use for the analysis of EBV?
10 October 2014 2,009 4 View
I am performing a very straightforward continuous assay where I basically follow a decrease in absorbance which corresponds to the conversion of my substrate to its product. However, I am getting...
10 October 2014 6,103 12 View
I have been trying to make this peptide for a long time. it's a 12-mer that is basic in nature according to a peptide property calculator. After making the peptide on a PTI Tribute Synthesizer, I...
10 October 2014 5,735 7 View
I have used 10 genomic DNA and a 650 bp probe validated to giving PCR amplification with the same DNA. I am following the standard Sambrook protocol for making probe and standard manual procedure...
10 October 2014 1,535 14 View
We obtained change in iNOS expression through Reverse trascriptase PCR, but failed to get change in fold expression through real time PCR. The CT value obtained was quite High 32-33 and almost...
10 October 2014 723 2 View
protein extraction
09 October 2014 7,180 2 View
08 October 2014 1,094 6 View
08 October 2014 6,836 2 View
I am cloning a gene into a vector that has the reporter gene LacZ, but I am getting some background and almost no clones. If I use another enzyme, to cut the reporter gene LacZ, would it be a good...
08 October 2014 2,872 1 View
07 October 2014 8,642 11 View
06 October 2014 1,767 4 View