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Questions related to Protein Purification Techniques
Hi guys. It's my first time doing RNA extractions. I'm trying to extract from soybean seed using the Promega kit. I think I've managed to get some RNA but judging from gel pic the quality is not...
07 July 2016 1,436 4 View
I am getting very intense non-specific bands in western blot, but very faint band of protein of interest. What should i do? please see the image and suggest.
07 July 2016 8,195 15 View
I have a kind of agarose-based scaffold and the size of that is like a well of 6 wells plate and I want to separate cells from scaffold for adding lysing solution and using a kit for extracting...
07 July 2016 1,567 9 View
hello we have in our lab Protein Electrophoresis and Blotting apparatus from biorad and I want to use it for RNA page electrophoresis. is it possible to use it? thanks for your help.
07 July 2016 2,190 3 View
I'm studied protein that produced from beauveria sp. and I'm doing SDS PAGE to see the profile (band, molecular weight)... is there anyone who already studying this and have the SDS PAGE result...
07 July 2016 3,827 1 View
As stated in the question. I would be grateful if anyone could guide me as to what I might be doing wrong with my Western Blot. I believe that it could have something to do with the transfer (too...
07 July 2016 5,432 6 View
I am aware of Streptavidin magnetic beads which can be used with biotinylated antibody in MACS. But I would like to know if anyone has tried isolating or depleting cells from heterogeneous...
06 June 2016 8,838 2 View
In a protein extraction, after the tissue grinding,suspension in TCA and centrifugation the pellet is obtained which is lyophilized to remove any traces of acetone. however in areas where a...
06 June 2016 5,033 4 View
I am trying to isolate CD34+ cells from cord blood and I have been getting 75% purity each time.So I am wondering have anyone tried using another column to elute them again. Will that help in...
06 June 2016 8,047 1 View
Hi Researchers, Currently I am doing 2DGE to identify differential protein expression (unknown) in plant tissues under various abiotic stress conditions. I am following phenol extraction...
06 June 2016 513 4 View
Hi everyone! I've been trying to do some simple genotyping for weeks. I've been painstakingly following the procedure that has worked for others in my lab, and have tried changes such as changing...
06 June 2016 6,195 7 View
I am using fresh-frozen tissue from P301L tau mice. I'm homogenizing 1 hemi in 1mL diluted RIPA (with dH20) with protease/phosphatase inhibitors. After centrifuging I take the supernatants and run...
06 June 2016 6,329 6 View
I'm trying to express membrane protein gene in pACMV-tetO Flag taged one. I did a transient expression in HeK293 Gent- but when I did a western blotting (without purification) only the cell lysate...
06 June 2016 4,957 8 View
Hello I extracted Chlamydomonas genomic DNA. and i checked gel electrophoresis. But i can't find genomic DNA in gel. and it remained smear. i didn't treated RNase. Can you check my protocol? 1)...
06 June 2016 1,107 3 View
I use 24mm diameter transwell(0.4μm) to culture Caco-2 cells. I want to detect expression of protein after cells exposed to cytokine. there are three methods first is directly put the lysis...
06 June 2016 5,464 4 View
Hi everyone, I've been testing 1:250 sirtuin ab (santa Cruz h300) and 1:1000 secondary ab. I think the problems isn't related to my samples since other proteins results were excellent including...
06 June 2016 2,717 13 View
I intend to separate ribosomal and non-ribosomal fractions of cytosol using 20% sucrose cushion. I have already run the cell lysate on top of the 20% sucrose solution for 2 hrs at 1,39,000 g. I...
06 June 2016 1,295 2 View
I am facing some problem regarding the residual phenol contamination in my DNA samples. I use phenol-chloroform-isoamyl alcohol mix to remove proteins. The 260/280 ratio suggests the presence of...
06 June 2016 8,872 9 View
Dear all, I try to detect possible mutations within my DNA. For this, I use the Surveyor Nuclease S Assay (Cel-I-Assay). After heteroduplex Formation andTreatment with Surveyor Nuclease, I don´t...
06 June 2016 8,163 3 View
HI, I am producing my recombination protein in Fetal Dog kidney cell. Although, It is a transmembrane protein and i am only producing protein for the ectodomain, thus my protein is secreted in...
06 June 2016 4,352 4 View
hello,everyone! I want to identify that Schwann cells which isolate from postnatal 1-4day Rats is differentiated when culture in differentiation conditions. By western blotting, I got bands of...
06 June 2016 6,871 5 View
My dear all, I purified trx tagged protein, but to cleave the tag I has to use enterokinase which works in the Tris buffer. My protein was in phosphate buffer.how can I transfer my desired protein...
06 June 2016 2,998 5 View
I'm doing SDS PAGE and the result show that precipicated protein with ammonium sulfate have a thicker band than the crude extract... is there any possibility of that happening? in my opininion,...
06 June 2016 451 6 View
I want to purify my protein which is clone in pET22b with a frame with pelB. my protein is small (11 kDa), I 'm searching a good protocol for extraction of my protein from periplasmic space. My...
06 June 2016 1,740 7 View