How can i find catalytic domain for specific protein kinase using cDNA ?

07 August 2016 8,217 0 View

Transfection should I grow the cells in Media without antibiotic and FBS ?

Transfection should i grow the cells in Media without antibiotic and FBS ?

05 June 2015 9,263 5 View

What should i expect on my SDS gel after analyzing my immunoprecipitation experiment?

05 June 2015 2,941 0 View

How can the cell attach to cell culture plate?

How can the cell attach to cell culture plate? I know there are two reasons. First, electrostatic reaction second, extracellular matrix but need more explanation.

03 April 2015 6,757 4 View

Can anyone suggest why the growth of my cell culture of TNBC MBA-MD-436 is slow?

03 April 2015 4,256 3 View

Need help I just start my first cell culture experiment .what's rounded shape cells means in cell culture during the growing in media no trypsin ?

03 April 2015 2,174 7 View

Can anyone please help me to find information about these 6 cell lines ?

kuramochi -ovcar - ovarsha- HEYA8- SKVO3- OVAMANA Please help me to learn more about these cell lines.

10 November 2014 624 2 View

Why am I getting a very dark high background in WB Kifc1 antibody?

I'm working now on western blot and trying to detect Kifc1, but it gives me a dark background and I washed for 2 hrs and used 5% BSA-TBST blocking. The same problem of high background is very dark.

10 November 2014 281 5 View

What's the importance of making standard curve for protein estimation ?

i do it but i can't understand the mechanism and how we can estimation our protein using Bradford soln ?

10 November 2014 4,244 4 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Which is a lucid standard comprehensive book to guide on psychological scale construction and its standardization?

02 March 2021 7,082 3 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

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

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

How to evaluate unsupervised clustering metrics when some elements do not have a cluster label in the result?

I run algorithms like HDBSCAN and MeanShift, and compare them with K-means and GaussianMixture. The first two algorithms do not return cluster labels for some elements in the dataset. My question...

27 February 2021 428 3 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