How to create a diagram/schematic of protein-protein interfaces?

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What should my PVDF membrane look like post transfer during a Western Blot?

When I perform western blot experiments I often get inconsistent results post transfer. What I mean is that when I look at my PVDF membrane after the transfer step I can often see the molecular...

08 September 2018 4,990 4 View

Why does my plasmid run heavy on an Agarose gel?

To test if my double digest of the pTriex-3 vector is working I ran my 'uncut plasmid', 'double digested plasmid' and 'enzyme 1 cut plasmid' and 'enzyme 2 cut plasmid' on an agarose gel. Double...

02 March 2018 4,293 2 View

Have my BL21 cells gained antibiotic resistance?

My BL21 cells when grown on an Ampicillin agar plate, completely cover the plate in a lawn as fast as 5 hours. They do this regardless of whether I transform them or not. A plate treated with no...

01 February 2018 384 5 View

How did my insert vanish from my plasmid?

Short version: My plasmid contained insert but later after a second transformation into a second cell line the insert vanished and does not show up on PCR or sequencing Long version: I used...

01 February 2018 6,758 9 View

How can I delete a large DNA fragment from my gene of interest using PCR techniques?

I have a gene of interest approximately 2000 bp long coding for a protein approx. 670 amino acids in length. I wish to excise one of the proteins domains from the gene. To do so I need to remove...

08 September 2017 2,155 6 View

What is the detection limit for a species (in time) of absorbance spectroscopy?

How long must a species last for, for it be detectable through typical absorbance spectroscopy. Say I have an enzyme with a turnover number of 300. Each reaction would take only 3.3 milli seconds....

07 August 2016 1,213 4 View

Do cumulitive effects on specific activity indicate a concerted mechanism?

I am looking at an enzymes reaction mechanism. I have investigated 2 mutations. both mutations individually reduce the enzymes catalytic activity. The 2 mutations combined render any product...

03 April 2016 2,818 10 View

How to determine the pKa of Glutathione (GSH) bound to proteins?

https://www.ncbi.nlm.nih.gov/pubmed/2742854I am following the method of Graminski in the paper above. Whether GSH is protonated or not may be followed with absorbance spectroscopy at 239 nm. To...

31 December 2015 3,812 4 View

Can you relate the energy of enzymatic steps to Kcat?

If I know that the free energy barrier of the first step of a enzymatic reaction has increased (And I know by how much) but the other steps remain unchanged may I infer from this any kind of...

31 December 2015 3,436 7 View

Can anyone help with problems with non linear elution pH gradients during Chromatofocusing?

I am using GE healthcare poly-buffer 96 and PBE 94. I use a 0.025 M ethanolamine start buffer pH 9.4 and an elution buffer of PB 96 pH 6.0. I take care to degas all my buffers, keep the...

06 July 2014 2,759 2 View

How can I prepare virus for a TEM or SEM imaging?

I have virus (viral hemorrhagic septicemia virus) in suspension and the experiment will not involve cells. What level of TCID50 is preferred?

11 August 2024 3,115 1 View

Which Scopus Journal provides the most affordable fees?

"PUBLISHING IN A SCOPUS JOURNAL" Researchers are now at a cross road. The critical need to publish in a Scopus or ISI, etc journal is ever vital. Journal Publication fees must be submitted....

10 August 2024 8,621 1 View

Seeking Advice on Viability and Execution of Undergraduate Thesis Topic?

Hello everyone, I am currently developing a thesis proposal and would appreciate your input on its viability and how to effectively carry it out. My proposed topic is: "Does the perceived threat...

10 August 2024 8,992 0 View

Who will be moral responsible for the death of thousands of people in the event of an earthquake?

Who will bear moral responsibility for the deaths of thousands of people in the event of an earthquake? Weeks and months remain before the onset of strong earthquakes that bring death to...

08 August 2024 6,134 12 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

Are there any instruments for studying time similar to the way it is in space?

There are a huge number of methods for studying objects in space, according to the senses (and not only). Mechanical, thermal, optical, acoustic, electrical, magnetic, based on particle beams,...

06 August 2024 7,102 0 View

Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...

06 August 2024 1,495 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Please explain how the plastic input value should be considered from the true stress-strain curve for the bilinear elastoplastic material model ?

I am working on Abaqus/Explicit(Quasistatic ) for the deformation of the auxetic structure model. Please explain how the plastic input value should be considered from the true stress-strain curve...

05 August 2024 454 3 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View