Are co-expressed proteins always seen as 2 bands on SDS-PAGE?

03 April 2015 8,584 7 View

What is the best method to purify basic proteins with pi 7.8?

03 April 2015 1,920 6 View

How can I increase the binding affinity of GST-tagged protein to GST sepharose column?

02 March 2015 5,281 5 View

How does the heparin column work for DNA bound proteins?

Hi guys, My protein is nuclear human protein and I am purifying recombinant protein from bacterial culture. when I performed ion exchange chromatography, my recovery was nil. thinking it is bound...

02 March 2015 4,341 9 View

Why is the zero'th time deacetylation reaction showing higher florescence intensity than 1hour reaction?

31 December 2014 1,904 1 View

Why is my protein acting differently at the same pH?

I am isolating a protein in soluble form from Bacterial systems. I have done cation ion exchange chromatography(Q column) at the pH 8.2 using Tris Cl and HEPES buffer. Protein is binding to the...

09 October 2014 9,222 10 View

Why is my protein getting precipitated, even at low concentrations?

I am purifying a recombinant protein (host Rosetta) with his tag. I am getting reasonably good yield (4-5 mg/liter of culture). However after affinity purification using Ni-NTA column, during...

05 June 2014 6,240 5 View

Can anyone help me figure out why I am losing protein during ion exchange chromatography?

I am purifying a protein from a bacterial system and I was able to successfully purify the protein through an affinity column. My protein has a pI of 5.9 and it is stable at higher pH. After...

05 June 2014 5,875 6 View

How much of Dnase can be added and what buffer can be used for cell lysis during protein extraction?

I am isolating protein from more volume of culture. However I am unaware how much dnase to use and also what buffer to use for its activity, if I am dissolving the pellet in 30ml of lysis buffer...

06 July 2013 4,166 5 View

More proteins are going to inclusion bodies, so how does one get a soluble protein?

I am trying to express a recombinant protein in E.coli, however the problem is with its solubility. I am inducing (0.5mM IPTG) the cells at 0.7 OD and incubating at 15 degree temperature. Most...

05 June 2013 5,457 14 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 View

Do you know good and user-friendly programs to measure plant quantitative traits?

For my research i will need to measure plant quantitative traits (especially leaves area and roots length, but would be nice to add some more information). I recently discovered...

01 March 2021 5,035 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

How to accurately quantify chitin?

Hi all, I have been working on the quantification of chitin in insect samples and feed for a while but I can notice that the quantification methods (e.g. exposure to HCl, NaOH, and weighing sample...

01 March 2021 1,843 2 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View