Do I need to strip the PVDF membrane before staining it with colloidal gold?

Dear Colleagues, After running Western blot on PVDF membrane and detection using ECL, I would like to stain my PVDF with colloidal gold to be able to allign the ECL image with total proteins on...

02 March 2021 7,829 3 View

Shall I skip the measurement model in SmartPLS?

Hello everyone, I had run a mediation model in SmartPLS using questionnaires that were already validated among my statistical population. A professor who edited my paper -though she is not...

01 March 2021 3,052 3 View

Have anyone checked before Cyclic voltammograms of intial ITO substrate and E-teck?

As a beginner in electrochemistry, I tried to obtain cyclic voltamograms of commercial anatase powder deposited on an ITO substrate or on Eteck. After several attempts, I noticed that my cyclic...

23 February 2021 3,175 3 View

Is it okay to mention you distributed your qualitrics study link through social media sites?

Just want to be clear and know it is okay to say I have distributed the qualitrics link through social media such as Facebook, Instagram and Twitter to ensure my target audience is reached in my...

21 February 2021 5,143 2 View

Why samples do not run in resolving gel?

Hello everybody, I have a problem with SDS-PAGE and I do not know why my samples stop at the top of the resolving gel and they do not run through it. The percentage of gel is 15% and the size of...

14 February 2021 9,116 6 View

Do you know of an informal caregiver of a person who has recently received a bone marrow transplant?

We are conducting a pilot study of an intervention for informal caregivers of persons who have recently received a bone marrow transplant in the US. If you know of anyone who might be interested,...

13 February 2021 9,174 1 View

Is it possible that the single-pass absorption exceeds 50%?

Dear researchers It's well-known that the maximum light absorption of any structure can not be greater than 50%. Is it possible that the single-pass absorption exceeds 50%? How?

17 January 2021 2,335 7 View

Most appropriate fit of a standard curve in an enzym-linked lectin assay?

Dear all, I have a question regarding the most appropriate fit of a standard curve in an enzyme-linked lectin assay set-up to quantify the amount of mucin secretion in conjunctival explant...

03 January 2021 1,010 6 View

How do I access Domestic Violence Attitude Scales or find them for free?

Hi, I'm a postgraduate student currently starting up a research project on attitudes towards domestic violence (DV). The study will aim to compare cultural differences between attitudes and...

31 December 2020 781 3 View

A good protocol for protein purification by AKTA?

Hi everybody, Does anyone have a good protocol for protein purification using AKTA? I have tried to purify two proteins but after several attempts it was not successful. So, I really appreciate...

28 December 2020 5,220 15 View

Looking for Materials with high thermal expansion coefficient and high temperature resistance?

03 March 2021 8,272 1 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View