Can any one send me some articles related to the role of MPS1 kinase in CHK2 actvation?

07 August 2016 4,368 0 View

What is appropriate incubation time for primary antibody in western blot analysis?

Primary antibodies, Incubation time, western blotting, ELISA, SDS-PAGE

07 August 2016 6,517 22 View

For phosphospecific kinase proetin how much time incubation (45') for secondary antibody is good for perfect result and washing time after incubation?

western BLOTTING, SECONDARY ANTIBODIES, PBS, Incubation time, SDS-PAGE

06 July 2016 9,545 2 View

Can blotting effect if some amount of SDS-Buffer present in transfer buffer solution?

Can blotting effect if some amount of SDS-Buffer present in transfer buffer solution?

06 July 2016 1,066 5 View

I am using BSA for blocking membrane for Phosphospecific antibodies but getting much background, could any one help me how to overcome this problem?

I am using BSA for blocking membrane for Phosphospecific antibodies but getting much background, could any one help me how to overcome this problem?

06 July 2016 2,109 4 View

I am working on CHK2 kinase protein and every times after western blot getting blackground stain also, washed 3 times10 minutes each,what is problem ?

western blotting, SDS-PAGE, CHK2, CHK1, Washing buffer, PBS, TBS

06 July 2016 9,108 8 View

For phosphospecific antibodies why milk blocking buffer is not good?

06 July 2016 5,507 1 View

Effect of ethanol on MCF-7 breast cancer cell line?

03 April 2016 5,196 2 View

Can any one please send me rad 9b gene related research article??

02 March 2016 3,858 0 View

What is main difference between Ct and Cq value? When do we use Ct value and when Cq In Real Time PCR?

11 December 2015 612 13 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

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

Anyone that can help with Maxquant TMT16plex which is missing?

Hi I am very new to proteomics analysis. I have got raw files from core facility running thermofisher TMT 16plex 4 hours. I cannot find on maxquant the option TMT16Plex there is only up to 11...

25 February 2021 1,675 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

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View