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Questions related to Protein Purification Techniques
I am trying to isolate the enzyme 5a-reductase from my PC-3 (Prostate Cancer) cell line. Does anyone have any advice to give concerning this extraction? For your kind advice, please. Thank...
01 January 2016 1,753 6 View
I am trying to extract matrix metalloproteinase-2 (MMP-2) protein from the cell. At first, I just used the common protocol (by RIPA kit) for whole cell proteins, but it seemed to be a wrong...
01 January 2016 9,225 1 View
I would like to quantify unknown protein concentration of plant seeds based on BSA at 540 nm
01 January 2016 9,223 11 View
Hi all, I am denaturing and refolding a protein complex containing disulfide bonds. After size exclusion and ion exchange, we're able to isolate what we think is correctly folded protein. I ran...
12 December 2015 4,887 4 View
Could you provide me with a protocol for best method of HA pulldown?
12 December 2015 5,852 5 View
Peptides are about 6.8, 3, 2.9 and 2.7 KDa and I use Tris-Tricine SDS-PAGE (16.5% and 18%) but gel resolution is not clear. so I can′t see 5 and 3.4 KDa bands of marker. could anyone help me with...
12 December 2015 1,483 7 View
Good day group! I have to purify a recombinant protein coupled to his-tag, wich is expressed in yeast. In my laboratory it is being evaluated as a method of analysis the "Y-PER Yeast Proteins...
12 December 2015 4,875 2 View
Hello, I am currently using LDH and bounding to toyo pearl chromatographic particles. I know I can determine the concentration of the protein in a buffer solution using the Bradford Assay Method....
12 December 2015 8,671 7 View
Showkat Ahmad How can we express the viral protein that is not complete ORF (i.e. with out start or stop codon) but a part of one complete gene and becomes functional protein after Post...
12 December 2015 6,187 4 View
I like to use Ion Exchange Chromatography for enzyme purification. I got an article where they used DEAE-cellulose 52 column. How can I use this and what instruments/apparatus I need to use this?...
12 December 2015 970 11 View
Hey, I'm trying to strip a dot blot without denaturing the protein in it's native conformation. Does anyone have any recipes for this?
12 December 2015 7,559 3 View
the proteins should be isolated and purified from the bacterial culture (B. Cereus)
12 December 2015 5,814 7 View
What are the drawbacks of Autoinduction of bacterial culture for protein purification? What about the stability of the protein expressed by such methods. Does anyone know about the report that...
12 December 2015 8,296 4 View
after setup crystallization screening, crystal were grown overnight in needle form. at this condition 5% Tacsimate 7.0 0.1M HEPES 7.0 10% PEG5K MME, but after 48 hours were disappeared, i have...
12 December 2015 3,190 3 View
I have an in-house raised antibody against my protein of interest which I have purified from the serum by the caprylic acid method and further confirmed on a sds gel. I wanted to use it for...
12 December 2015 2,718 7 View
i have precipitated protein using ethanol........do i need to concentrate the protein before i start chromatography techniques......... if yes then what centrifuge membrane tubes should i use for...
12 December 2015 3,526 6 View
I expressed the recombinant protein and purified with Ni-NTA in pure form (Single band). I set up the dialysis to remove urea and imidazole from the purified protein sample (10ml). After dialysis,...
12 December 2015 4,344 9 View
Hi, I´m trying to extract protein from mouse lung for WB, using TR3 lysis buffer consisting of glycerol, SDS and Na2HPO4, but I´m facing a problem at the end step when I try to take the...
12 December 2015 6,095 7 View
The ultrafiltration/diafiltration is the last step for protein purification. During the ultrafiltration/diafiltration of protein solution, i found TWEEN 80 was adsorbed by membrane. I want to add...
12 December 2015 9,634 4 View
I'm working on a membrane protein. For now I have gotten many difference type crystal. I find the most beautiful crystal has the most bad diffraction (< 20A）while others (6~15A). But I don't...
12 December 2015 9,073 5 View
Has anyone had any successful experience generating polyclonal antibody (in rabbit)? If so, could you please share your experience and the company that you decided to generate the antibody...
12 December 2015 4,227 2 View
I need to prepare a modified SDS-PAGE to run a sample with modification. It is easy to lose its modification for my protein in 6.8
12 December 2015 4,238 5 View
I have cloned my gene of interest in pcDNA3-CMV-APEX-NLS and this construct was used to transfect HEK293T cells.pcDNA3-CMV-APEX-NLS was used as a control during transfection.I have performed...
12 December 2015 3,462 3 View
the protein is a transcription factor subunit of AP1, the sample was treated with B-ME and boiled at 95 degrees for 5 min, ran on SDS-PAGE, could it still be in a dimer after all this?
12 December 2015 6,318 5 View