Why are my western blot bands are think and faint?

Hi, my western blot bands are appearing at the correct molecular weight but are very thin and faint, making it difficult to distinguish difference in expression for shrna samples. Conditions...

04 January 2021 9,204 4 View

Nuclear protein extraction protocol from cells?

What is the most suitable protocol to extract nuclear protein from cancer cell lines?

29 October 2019 4,748 3 View

How to purify GABA from microbes?

I am working on neurotransmitter from microbes .

26 January 2018 651 1 View

What are the possible impacts of implementing multidimensional information modelling in construction ?

08 May 2016 5,507 1 View

How do I connect a 2 Wire strain gauge to QuantumX MX1615B without adding temperature compensation?

I need to connect an HBM K-LY 4/120 strain gauge(Linear 2 wire) to HBM QuantumX MX1615B DAQ system. This will be mainly done for the calibration of a custom built amplifier circuit. Although...

27 December 2015 8,014 2 View

Does there exist periodic Gaussian function?

Is there any way of producing periodic Gaussian pulses such that the bell shape repeats itself with a defined period. Does there exist any solution for some differential equation which resembles...

26 May 2015 2,986 13 View

Can anyone help with co-transfection of miR overexpression with its target?

In order to see the functional activity of my miR-over expressing constructs, I did a co-transfection of the over-expressing plasmid with its target reporter. I could hardly see much difference...

10 June 2014 5,725 5 View

Why should additional NaCl be added for nuclear protein extraction (nfkb isolation)?

I am trying to isolate a nuclear protein NF-kB from raw cells in order to do a western blot. I have searched some protocol for nuclear isolation and finally came up with one. But in that...

11 April 2014 2,153 3 View

Malware image data set for research purpose

I am working on malware analysis. I know that publically available data set Malimg. I woulde like to know is there any data set publically available for research purpose.

01 January 1970 7,440 11 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

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 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

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 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

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View