When collecting data to conduct CFA, can I collect an expanded set of items and then limit the CFA to the reduced set of items from the EFA results?

I have been working of developing a new measurement tool. I have followed all the standard scale development steps / best practices and have a reduced set of items (from 31 down to 22 items via...

02 September 2019 2,598 3 View

Why am I getting increased transformation efficiency in yeast transformation protocol when I have changed nothing ?

I have been working is S.cerevisiae for about 2 years. Up until about 8 months ago, I would have successful plasmid transformations with about 30-100 colonies per plate depending on the size of...

28 January 2018 1,321 3 View

Tau Protein Western Blot Analysis in ImageJ

Can anyone explain how to analyze western blots that contain multiple bands for a target protein in ImageJ? This image is for Tau 5 (total tau) but this also happens for pSer396. This is...

01 July 2017 8,874 6 View

How much folate is provided to a plant from a microbe symbiont?

I work with methylotrophic bacterial plant symbionts. I know that the bacterial symbionts produce and release folate into the soil that the plant will take up. Does anyone know about how much of...

30 August 2016 9,660 1 View

Live decapitation preferable to isoflurane or CO2 asphyxiation?

I was speaking with our University's vet and she mentioned that live decap is the better route to take if you're planning on doing any histology after death. Does anyone have any further...

19 July 2016 4,669 6 View

Does anyone know whether hTERT-RPE1 cells express bestrophin-1 protein? Do you know whether they polarise or differentiate?

We are trying to find an RPE cell line that expresses bestrophin-1 protein. We have established that hTERT-RPE1 and ARPE-19 express the RNA however do not appear to produce the protein. We would...

06 October 2015 712 2 View

Where does my AU Rich Element (ARE) region begin?

Hi, I'm working on a suspected ARE region in a 3'UTR of a transcription factor. Using the ARED website we've found that there are two "AUUUA" pentamers in our 3'UTR that are close together that...

15 February 2015 9,005 4 View

Is anyone familiar with nested PCR?

I started running nested pcr this week using Extract N Amp. This is my first time doing this, and after the second reaction, my water negative showed positive bands. I will look into contamination...

25 September 2014 7,779 3 View

How to select substrate concentration to determine the inhibitor behavior as competitive or non-competitive?

I already calculated the IC50 value of the inhibitor against the protease enzyme. Now I need to find this inhibitor competitive non-competitive. I decided to perform the Lineweaver Burk plot, but...

21 January 2021 8,378 3 View

Best purification method for recombinant protein in BL21?

Hi I need to purify a recombinant protein (IL6 21kDa). It's been produced in E.coli BL21 with a T7 expression vector, but it does not have a tag as yet. I was thinking of adding a his-tag and...

09 December 2020 2,200 8 View

How to handle milk clotting Issues during media preparation for Screening of Halotolerant Protease Producing Bacteria (with Skim Milk)?

We want to screen halotolerant protease producing bacteria. Since Skim milk (HiMedia) is available in our lab, therefore, we want to screen extracellular protease by this compound/ supplement. We...

04 December 2020 7,657 6 View

Can TEV protease cleave between glutamine and methionine efficiently?

Based on this paper The P1 ' specificity of tobacco etch virus protease ,TEV protease can cleave between the Gln and several amino-acids (besides Gly/Ser) with acceptable efficiency in its...

26 November 2020 5,987 4 View

I need equations to calculate the activity of the enzyme glutamanase and basic phosphatase in soil and plants?

I need the rates for calculating the mentioned enzymes after the laboratory work. I have trouble calculating and extracting the activity of the mentioned enzymes in soil and plants because of the...

03 November 2020 8,601 1 View

What is the best method to check interaction between a drug and a protein ?

Hi, I would like to know the best method to check the interaction between a proteine (cruzipain, cysteine protease) and a drug. It has been purposed to our project to evaluate protein-drug...

30 October 2020 7,209 3 View

Does 2-methoxyethanol reacts with air?

Hello, I'm trying to use 2-methoxyethanol/Cellosolve to dilute methylumbelliferyl phosphate for phosphatase activity analysis. However, we never used this solvent before. It came with a sure/safe...

27 October 2020 7,302 1 View

How to purify a recombinant protein from Ecoli to reach a cell culture purpose?

05 October 2020 502 7 View

Protease activity assay?

Hello, I would like to get help! I want to measure the protease activity of pancrease powder. I prepared 10mg/ml stock solution from the pancrease powder and I used 100ul from that and I added...

22 June 2020 1,609 3 View

Why my protein has lower molecular weight that expected?

Hi, I am trying to make a kind of recombinant protein (Flag-tagged) from HEK293T cells. But when I check the cell lysate by WB, the molecular weight is lower that expected. The molecular weight...

09 June 2020 6,414 13 View