When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

What is the best temperature to coat an ELISA plate?

Hello, I want to control the temperature of my ELISA plate during incubations to get consistent results. In general, what is the best temperature to coat the plate with antibody? I have been...

16 February 2021 2,377 7 View

SPSS - Merging files by ID numbers that contain letters?

Hi, I'm trying to merge files from a data base of 20 files, each containing about 4000 subjects. Unfortunately the ID numbers are actually a combination of numbers and letters (e.g., b1458219)....

15 February 2021 5,833 3 View

How would I go about getting an expert review on the interview questions for the dissertation?

Good Evening, My name is David Marshall and I am Doctor of Management Student at Colorado Technical University Online. I just recently received approval from the IRB to start data collection on...

08 February 2021 346 7 View

How can I quantify the color of a solution?

I know colorimetry has been the standard way by way of measuring the concentration of a particular compound (solute) in a colored solution (solvent or measure quantities of a particular compound...

08 February 2021 2,673 1 View

ACE2 Blotted as a double band?

Hello, I have recently run a western of Vero cells for ACE2. I detected ACE2 at the correct size, but it presents as a double band. The sample was prepared by lysing cells in 1X Laemmli...

07 February 2021 6,973 3 View

SARS-CoV-2 Spike Protein Western Bands?

Hello, I am western blotting SARS-CoV-2 spike protein from lentiviral using an antibody against S1. To prepare samples, I mixed concentrated lentivirus 1:1 with 2X Laemmli buffer and boiled...

07 February 2021 3,643 4 View

How does DMSO percentage affect ELISA?

Hello, In my sandwich-type ELISA which is testing for small molecule inhibition of a protein-protein interaction, it seems that DMSO percentage and dilution protocol are important. I...

31 January 2021 6,771 1 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

(western blot) can methanol damage my antibody signal?

I accidentally rinsed with methanol after adding my primary antibody to my membrane. I noticed after ~10 seconds and immediately replaced it with PBST (3x15 min washes). Will my antibody signal be...

12 February 2021 5,824 3 View

What is the reason, we got both test and control band on LFA in negative sample?

I am working on Lateral flow assay for virus detection by carbon conjugated polyclonal antibody. We got a high intense band in both test and control line on LFA in negative sample used as buffer....

11 February 2021 6,558 2 View

How do I prepare and store insect samples for JH measurements?

I need to preserve termite samples for future quantification of juvenile hormone in their bodies. Or extract the Juvenile Hormone and keep that samples stored for future quantification. I may need...

09 February 2021 1,995 1 View

Background and spots in Western Blot membrane?

Do you know why my membrane looks like this? I did this western for Anti-hnRNP D/AUF1 antibody ab50692, as primary antibody in 1:1000 overnight at 4c and Anti-rabbit IgG HRP linked antibody...

07 February 2021 5,591 4 View

During an anti-His capture ELISA has anyone had issues with histidine residues from secondary antibodies non-specifically binding?

Hi, Hope you can help. Currently been running some sandwich ELISA assays where I coat with anti-His, block and then capture His-tag protein and then sera and then anti-bovine monoclonal...

07 February 2021 7,101 1 View

How do I keep my prefixed (PFA) frozen tissue sections from lifting off the slide?

I'm trying to stain mouse lung tissue IHC-IF I fix in 4% PFA overnight immediately upon harvesting tissues Then dehydrate/cryoprotect in sucrose gradients 15% to 30% Then freeze in OCT media...

06 February 2021 2,336 3 View

Can anyone suggest me broad host range fluorescence expression vectors for Gram-positive bacteria ?

Hi everyone, I am looking for fluorescence expression vectors for expression in gram-positive bacteria present in the skin microbiome. I need to tag them with a fluorescence marker and observe...

06 February 2021 2,566 4 View

Antibody dilution for IHQ in OCT frozen tissue is more similar to IHC in Paraffin-embedded samples or to Immunocytochemistry?

31 January 2021 8,678 3 View