What is 5X binding buffer?

Kyungsun Ha @Kyungsun_Ha

01 January 2019 0 5K Report

I saw 5X bingind buffer maded by;

100mM HEPES, pH 7.6
5mM EDTA
50mM ammonium sulfate
5mM DTT
1% Tween-20
150mM potassium chloride

which reaction helps this buffer? I don't consider to this buffer participates in which experiment by name.

Badges
Science method

More Kyungsun Ha's questions See All

In CHURCH buffer, what components do role at southern blot?

Greetings. I cannot find about each component's role of CHURCH buffer at southern blotting(detail, pre-hybridization). I did DNA microarray but never have ever southern blot. In microarray,...

31 December 2018 885 0 View

In alkaline transfer buffer, why put NaCl to alkaline transfer buffer?

Greetings. Before question, I never did southern/nothern blot. (eastern too) During search buffer, I found Alkaline transfer buffer and knew it uses southern blot(alkaline method). This buffer...

31 December 2018 8,961 0 View

In SDS-PAGE, how Tris-glycine buffer without SDS works?

Greetings. Here is sunday... In tris-glycine buffer, there are either include 1% SDS or not. I used former but many seller selling both Tris-glycine with SDS and without SDS. SDS is set charge...

31 December 2018 1,224 0 View

Why we don't have to use HCL/NaOH during make Trice-glycine buffer?

Greetings. I heard about when make buffer include Tris, set pH using concentrated hydrochloric acid(37%). but SDS-PAGE, loading buffer(10X), Tris-glycine, what I heard set pH for 8.3, use only...

31 December 2018 3,971 3 View

How can I measure tissues for RNA preparation without RNA dergadation?

Greetings. I extracted RNA at mouse liver and human blood, but degradation occurs. I think, at mouse liver, too hard to measuring 50mg liver(I am newbie for animal tissue preparation) so spend...

31 December 2018 4,671 0 View

What's the problem in gel electriphoresis?

Greetings. In image... left:ladder(start with 750bp) middle: PCR product with exonuclease III treated right(3rd lane): PCR product without exonuclease III treated there's a problem. PCR...

10 November 2017 9,858 12 View

Why stability test of PEG gel run at -20 degree?

I read a paper about PEG gel, . in this paper, supplymentary fig 10 is storage stability test. This particle maded by polyethylene glycol and store in 4 degree, but storage test is run at -20...

09 October 2017 2,833 0 View

Among the features of microparticles, what should be compared experimentally?

Greetings. Experiments using microparticles are in progress. There are particles with multiple compositions and features, advisors want to compare the features of this particle finely, but I can...

08 September 2017 535 0 View

What is Dunn's multiple comparison test?

Guten morgen.(means 'Good mornong' in Deutsch) Recently I read a paper entitled "Circulating Tumor DNA Mutation Profiling by Targeted Next Generation Sequencing Provides Guidance for Personalized...

08 September 2017 5,063 1 View

How can I add sodium chloride in 1X PBS to 450mM?

Oh, greetings. I plan to experiment for 'stringency of DNA'. so I add some sodium chlodire to PBS. and make 1X PBS(450mM NaCl). It same to concentration of NaCl in 3X SSC. But in 1X PBS, I buy...

08 September 2017 8,717 0 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 do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 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

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 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

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