How to store gel for MS analysis?

Please suggest, how to store a SDS-PAGE gel, after colloidal coomassie staining? I have to do LC/MS analysis, but instrument is not available for few days. can i store the gel? if yes how long? in...

05 June 2017 1,414 5 View

How we can analyze electron density map of cryo EM structure?

11 December 2016 1,965 4 View

Why the protein is not denaturing in SDS PAGE?

I am working with a protein of approx 35kDa, in SDS PAGE the band is coming at approx 72 kDA. If it is in Dimeric form then why it is not coming as monomeric form. Laoding buffer contains 5% BME,...

09 October 2016 8,592 8 View

Had any body done Circular Dichroism studies of ATP bound proteins?

I want to study conformational / structural change in the protein after ATP binding ( I am working with a yeast protein that shows ATPase activity) so I want to see and compare the binding effect...

08 September 2016 4,347 4 View

Does the ratio between CD spectra at 208 and 222 nM Matters?

For Circular dichroism studies, we know that if the negative elipticity at 208 and 222 are more then it would be alpha helical protein. but if it is more at 208nm or more at 222 nm, what does it...

06 July 2016 2,196 4 View

What is the minimum concentration of protein required to get Circular dichroism data ?

06 July 2016 8,317 5 View

Can any one tell me why my RT (reverse transcription) is not working?

I have RNA, isolated from Yeast cells (By TRI (SIGMA) reagent), quality is good after Dnase-1 treatment (on Gel), but cDNA is not formed. MMLV RT is working fine for other strains (RNA samples)....

03 April 2016 9,529 9 View

Why does the protein not bind to PVDF/ MCE membranes? Did anybody face this problem in filter binding?

I am doing RNA-Protein interaction exp. I want to see binding by filter binding assay, I used Millipore PVDF 96-well filter plates, with vacuum manifold, but could not see binding, I also tried...

07 August 2015 2,391 4 View

What are the importance of these compounds in synthesis of drugs....1-Indanol , Methyl-4 pyridine methanol , 1-Naphthyl)ethanol ?

What are the possible applications of these compounds..1-Indanol , Methyl-4 pyridine methanol , 1-Naphthyl)ethanol?

06 July 2015 8,974 0 View

What could be the reason for protein degradation during sample preparation?

During sample preparation for SDS-PAGE, some of my samples are precipitating metimes. They give smear like band on gel, what could be the reason? Heating at 95 for 5min and then cooling on ice for...

01 February 2015 6,885 7 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 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