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05 June 2019 2,765 4 View

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08 September 2018 3,685 4 View

How much medium is required to deactivate 1 mL trypsin?

As I know from researchgate question & answer session; "Split ratio depends on the volume of trypsin + medium to deactivate trypsin + cell" In this regards, my question is this statement correct?...

06 July 2018 1,666 0 View

Either anionic or non-ionic detergent is suitable for preparing lysis buffer?

I am presently planning to do western blot. In addition, I am working H9C2 cell lines. So, I need to prepare sample from cell line. Before to start, I have to prepare lysis buffer. In this...

05 June 2018 5,779 5 View

What are the shelf-life and optimal storage conditions for cell culture media?

I am currently working with H9C2 cell lines. I prepared the complete growth media by using following: 1. DMEM 2. 10% FBS and 3. 1% PS Finally, I know media (DMEM) by itself is generally stable for...

05 June 2018 9,877 4 View

What is the optimal storage condition of Sodium fluoride (NaF)?

In lysis buffer, Sodium fluoride (NaF) act as a serine/threonine phosphatases inhibitor. I found different concentration as well as different storage condition depending on its application over...

05 June 2018 2,763 4 View

How to prepare the Fixer solution?

I need a complete protocol to prepare fixer solution for X-ray film development. Note: For western blot Enhanced chemiluminescence (ECL) detection method.

04 May 2018 9,812 0 View

How to know the cell H9C2 is fully differentiated?

31 December 2017 5,637 5 View

Best method for superoxide dismutase measurement ?

I am presently working with H9C2 cell. Now, I want to quantify superoxide dismutase enzyme after exposure of hypoxic condition to H9C2 cells. So, which methods should I have to use?

11 December 2017 8,789 1 View

How can I choose best methodology to determine apoptosis?

In particularly, I want to know what criteria should I have to consider before selection of a apoptosis methodology? In addition, I am presently working on Ischemic reperfusion induced apoptosis...

11 December 2017 6,257 2 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

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

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

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

How optimize the Size Exclusion Chromatography?

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

Why are my bands weird in this Agarose gel?

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

What do I do with DLS peaks that are from the buffer?

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

High aspect ratio hexahedral mesh optimization solver?

I am interested in evaluating mesh optimization solvers that converge to high aspect ratio unstructured hexahedral grids. In other words, I am interested in a solver that optimizes warpage,...

26 February 2021 7,279 2 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

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

How to interpolate from a time-series database ?

Hi, I am having trouble with a problem, in the field of Optimal Control and the generation of optimal time-series. Let's consider a system, whose dynamics are represented by dx/dt =...

24 February 2021 9,105 2 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

Cloudy/high background western blot - can anyone help me?

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