Hi All,
I am planning to purify a His-tagged protein from HEK293T cells after overexpression. Could you please share your opinion regarding the right composition of the lysis buffer? I will be using FPLC ACTA.
Our lab is facing a difficult problem for almost a year. After thawing the cells are okay, but after 2nd or 3rd passage there are debris present between the cells. We use mycoplasma removed cells...
17 January 2021 5,629 6 View
Multi-objective optimization through chicken swarm optimization technique. Both maximization and minimization problem.
31 December 2020 2,299 4 View
I have used mosaic to new raster tool to join two aerial image from Landsat 8 of different path. However, raster files had color variation and after mosaic I got a straight line running from...
22 December 2020 4,640 4 View
How can we best quantify double-stained markers when comparing diseased and control? For example, if we want to stain one marker of smooth muscle cells and compare the expression in diseased and...
21 December 2020 7,389 1 View
Hello, I am expressing an integral membrane protein (MW 51kDa) in Hi5 insect cells. I initially purified the protein by Nickel-His, concentrated it and subjected it to Size Exclusion...
10 December 2020 2,781 5 View
I want to extract RNA from human breast milk. How should I store the milk sample in RNAlater? And after RNA extraction how can I store the RNA?
01 December 2020 7,183 4 View
The problem is that everytime getting the flavonoids content much higher than phenolics which may be, but in rare occasion. In phenolics, I am adding100 ul sample with 900 ul dH2O with...
29 September 2020 4,993 6 View
Hi All, Recently, I purified 8 proteins and stored at -20 with the elution buffer containing 20 mM Tris, 250 mM NaCl and 200 mM Imidazole. I autoclaved the buffer, so I do not know whether the...
25 September 2020 9,814 6 View
Aethalometers are used to estimate the light-absorbing aerosols or Black Carbon (BC) or Soot particles. There are widely used aethalometers which are measuring light absorption in multiple...
06 September 2020 1,050 3 View
I am trying to model magnetic target localization in MATLAB using those algorithms. So essentially what I want to output is the position vector of the target (x, y, z) and the magnetic moment...
28 August 2020 9,811 2 View
Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...
03 March 2021 1,499 2 View
Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...
03 March 2021 6,299 3 View
Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...
01 March 2021 2,215 2 View
Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.
01 March 2021 9,952 3 View
I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...
01 March 2021 9,015 2 View
I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...
26 February 2021 7,129 3 View
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
Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...
23 February 2021 6,266 3 View
Good morning everyone, Due to the pandemic, I conducted several online surveys. It is quite agile, not at all expensive, but there are several problems: the sample's representativeness and its...
21 February 2021 7,457 3 View
Hello everyone I would like to improve the buffer capacity of seawater (pH ~8) in order to dissolve as much CO2 as possible without dropping the pH below 7.5 (a range between 7 - 7.5 is...
17 February 2021 2,797 2 View