What is the typical workflow of mutant protein expression in mammalian cells?

I want to express some variants of human anion-exchanger transporter protein in K562 cell line using viral transfection method. What is the typical workflow for achieving this. No need to go into...

10 November 2019 5,708 3 View

Is it possible to extract protein from animal tissue by homogenizing in water only, without any lysis buffer or salt?

The objective is to simply quantify the protein by Bradford method. Is it possible to just homogenize the tissue in water, centrifuge it and analyse the supernatant with Bradford's reagent?

05 June 2018 2,224 5 View

How to convert a test tube assay to microplate assay?

I am trying to measure Ascorbic acid levels in tissue using Omaye et al protocol (Methods in Enzymology, Volume 62, 1979, Pages 3-11). This method is based on reduction of ferric chloride and...

31 December 2017 9,231 6 View

How to do statistical analysis of the enzyme kinetics parameters (Km and Vmax) among several treatment groups?

Lets say I measure kinetics of an enzyme under three toxic chemical exposure conditions. After fitting the enzyme velocity data using Michaelis Menten model, I am able to get the kinetic...

11 December 2016 4,098 9 View

GeNorm in NormqPCR package of R?

In the original geNorm VBA applet, the raw Ct values were required to be converted in the relative Ct values (by using PCR efficiency and lowest Ct among samples). Does 'selectHKs' function in...

08 September 2016 7,933 5 View

Are 'predicted' protein sequences useful for multiple alignment in order to draw conclusions about conserved domains?

I was wondering if predicted protein sequences, which sometimes come out of the 'Gnomon annotation program' of NCBI, can be used for multiple sequence alignment in common tools such as Clustal...

02 March 2016 2,379 1 View

What will be the impact of increased enzyme expression on its Vmax and Km?

Will increased enzyme expression lead to increase in Vmax ? Ideally, it should be. What is your opinion on this?

07 August 2015 5,988 12 View

How do I remove skin from small fishes?

Is there any specific protocol for removing the skin from small fishes (

06 July 2015 2,118 8 View

Is it possible to statistically compare two values if we do not have information on replicates?

E.g. Is it possible if we want to check statistical significance in the difference between two separately published LC50 for a particular chemical in two separate species? To elaborate lets take...

05 June 2015 4,721 18 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

How the parameters (such as Mw, Chemical bonds, polymer bland and so on) affect the mechanic property of polymer (such as storage, loss modulus) ?

Anyone can recommend any books on such topics? Thanks in advance.

02 March 2021 8,457 2 View

How to grow E.coli on the low salt LB agar plate with 50 or 100ug/ml of Blasticidin?

Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...

01 March 2021 2,027 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

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

How to make fractions of chloroform extract of plant, using seperating funnel?

28 February 2021 8,466 2 View

Can Ethovision store the generated data and video recordings directly to NAS system?

The size of data generated by Ethovision is huge. Having NAS system for data storage seems to be a good option, I am wondering whether Ethovision can directly store the data to the NAS system...

27 February 2021 6,350 2 View

Recalcitrant seed short-mid storage method?

Dear peer scientists, Ultradry storage is not suitable for recalcitrant seeds. However, theoretically, if we can stabilize the structure of the subcellular membrane, as well as glassify the...

25 February 2021 2,442 1 View

Can two EC50 values with different hill slopes be compared?

Hello, I fit a classical dose response curve Y=Bottom + (Top-Bottom)/(1+10^((LogEC-X)*HillSlope)) to my data (not related to inhibitors etc). The model I use is purely qualitative but it fits...

23 February 2021 8,155 5 View

How to calculate the fraction metabolized of a drug by an enzyme using clearance(ml/min) values of the metabolite formed?

I am trying to find the fraction metabolized of a drug by CYP and UGT enzymes. I am unable to get the values of fraction metabolized (Fm) directly from the literature. Can we calculate the Fm of a...

21 February 2021 5,379 2 View