We want to measure MCT1 and 4 on human muscle samples by the use of western blot. On which exact kDa appears the band? Is this a little higher than the expected 42 kDa (eg 49). Information is much appreciated.
I want to do L-Glutamine injections in mice to check mTORC1 activation in muscle. I cannot find dose-response or time-course studies in literature. Could anyone help me out?
31 December 2016 6,462 1 View
I was wondering if there was an antibody available yet (commercially) for phospho-sestrin2 (Thr 232, Ser249 or Ser279). According to the paper attached Sestrin2 should be able to get...
08 September 2016 7,032 2 View
Does anyone have good reference papers looking at the increase in leucine concentration after a standard meal (leucine gavage) in humans and mice?
09 October 2015 590 2 View
I've tried the novus #NB100-122, got a lot of (aspecific) bands.
06 July 2014 5,842 9 View
The venous samples have been taken in normal EDTA tubes, centrifuged and then stored in -80°C. They have not been DE-proteined by perchloric acid.
06 July 2014 8,178 1 View
Pharmacological, not siRNA or others.
06 July 2014 6,757 2 View
We recently put a couple of ML of TRIzol in plastic tubes. RNA extraction afterwards yielded contaminated RNA, indicated by a >2,4 260/280 ratio. Is it possible that the storage of TRIzol in...
03 April 2014 3,532 1 View
We are doing Western Blots with home made gels. The problem we are having since a couple of months is that the glass cover plates regularly break when gel-electrophoresis is finished. We use a...
07 August 2013 7,109 6 View
I'm looking for a method to generate membrane fractions (not crude membrane, but cell membrane fractions) out of frozen (-80°C) human muscle biopsies. I know it's usally done by sucrose gradient...
05 June 2013 1,448 1 View
I normally let the gel harden overnight, but is this necessary? Can I, for example, prepare the gel in the morning and load it with proteins in the afternoon?
03 April 2013 2,879 2 View
I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...
10 August 2024 7,480 3 View
I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...
06 August 2024 3,130 2 View
It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...
01 August 2024 4,673 4 View
Greetings. I’m currently running a nested pcr for giardia. My mastermix comprised of 3mM Mgcl2, 5unit of taq polymerase, 0.2uM of forward and reverse primers each respectively and 0.2mM of dNtp...
22 July 2024 9,761 5 View
I’m having difficulty achieving high RNA integrity in my samples. Although the 260/280 and 260/230 ratios are satisfactory after RNA extraction, the RNA samples show signs of degradation when...
22 July 2024 155 4 View
Sometimes I see the shadow like bands and its not true band. I want to know that what's the reason for it. I am using 2% gel for running genotyping samples I have uploaded the gel picture in both...
19 July 2024 148 6 View
Hi, I have lysate from human macrophages, and I wonder what will be the best way to check the NFKB activation in those cells. I was thinking p-p65 or p-ikkb./ Is that make sense?
18 July 2024 5,214 4 View
I have been running native page for FAM DNA substrate ( fluorescence samples) for protein DNA binding reaction. Binding is there but towards the end of the lane , I am loosing signals...
17 July 2024 6,213 4 View
I start with IDT ordered 124 bp ssDNA that is PCR amplified, then I use a QIAgen agarose gel extraction to get my exact product, and I'm trying to purify ~95bp RNA by denaturing PAGE to prevent...
16 July 2024 4,604 0 View
Having trouble detecting phosphorylated HSL. Any tips for how to prevent de-phosphorylation events or detect phosphorylation?
15 July 2024 2,055 3 View