I'm going to do Western Blot and I would like to know what is the best kit to use to extract the protein from adipocytes cell (3T3-L1)?
You may use M-PER (Mammalian-Protein Extraction Reagent) from thermo-pierce. Best...
MPER with Protease inhibitors works fine. I did in hepatic cells. Extraction was fine!!
I'm running Real-time PCR on STEPONEplus instrument (singleplex) which allows me to choose only one endogenous gene. However, can I run another endogenous gene in the same 96 well? I would also...
04 May 2013 2,393 2 View
dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...
03 March 2021 6,994 5 View
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
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
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
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
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
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
Warm greetings My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology (on Human papillomavirus). At this stage, I have almost completed a clinical sample Cervical swabs)...
25 February 2021 1,660 3 View
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
Warm greetings! My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology focusing on Human papillomavirus). At this stage, I have almost completed a clinical sample (Cervical swabs)...
25 February 2021 3,612 3 View