I want to elute using a pH gradient for a protein bound to Ni-NTA resin. What are the optimal solutions to create the gradient?

17 March 2022 0 415 Report

I am trying to purify a His bound protein off of a Ni-NTA resin and cannot find a protocol that says the chemical make up of the solutions used to create the gradient.

Badges
Science method

More Casey Callies's questions See All

Prevent evaporation in hypoxic incubator?

I have noticed massive media evaporation (>50% volume) from culture dishes after 72-96hr inside a Memmert ICO50 hypoxic incubator (Temp: 37C, humidity: 86%, CO2: 5%, O2: 1%). I've confirmed the...

29 April 2024 9,872 1 View

How does the stability of routine biochemical analytes in whole blood and plasma from lithium heparin gel tubes change during a 6-hour storage period?

The inquiry relates to a study conducted by Alkouri, Rana, et al. in which they investigated the stability of various biochemical analytes in whole blood and plasma samples stored for different...

15 April 2024 3,962 0 View

What are the preparations needed to be followed by the patient during the collection of their urine & how can microbiologist ensure that the patient?

The question is based on the article, “Tenets of Specimen Management in Diagnostic Microbiology” by Rajeshwar and Pathak. It aims to know how the microbiologist can help the patients do their part...

05 February 2024 9,935 3 View

How can I interpret this ITC result?

I am using isothermal titration calorimetry to measure the binding affinity and stoichiometry between a small molecular weight poly(amino acid) (3500 - 4500 Da) (SYRINGE) and high molecular weight...

02 August 2023 6,564 8 View

Can any one help me interpret the results of a restriction digest of a plasmid DNA the vector was pET22b(+) and the enzymes was Nde1 and EcoRI?

the cloning , transfornation and plasmid prep was done in the a series of lab sessions too . The pet22b(+) vector was combine with the ALDH gene . The digestion is done on the eluted DNA from the...

26 October 2022 2,660 1 View

Using Gen5 Cytation for Micro BCA Assay - How is a protocol set up with unknown dilutions?

I am trying to using the Gen5 Cytation by BioTek for protein quantification using a micro BCA Assay kit. I downloaded a BCA Assay Experiment (.xpt) File, and tried to use it but ran into some...

21 June 2022 650 3 View

Check that Repli-G MDA reaction worked before sending product for sequencing?

I want to check that my single-cell multiple-displacement amplification (MDA) reaction worked before I send the product for sequencing. From the Qiagen protocol, running a gel wouldn't be...

12 June 2021 6,317 2 View

Can I use soft vacuum filtration to harvest Ath seedlings from liquid media?

Hello, I'm growing different accessions of Arabidopsis thaliana and so far we have been harvesting them by pouring the media over a simple sieve, then putting them on paper towels for a minute...

18 February 2020 8,139 3 View

Are you considering genetic genealogy sources for YSNP and YSTR. GEDMATCH is being used daily now by US law enforcement but not YSNP & YSTR data?

With 10M testers with atDNA and public access via GEDMATCH, dozens of law enforcement agencies in the US are using this source with amazing results for cold cases and unidentified remains. YDNA...

23 January 2019 1,280 5 View

How to address and/or avoid Klebsiella contamination of primary human bronchial epithelial cells?

We have isolated primary human bronchial epithelial cells and frozen them down. After thawing and culturing them, and having identification done, it appears that we have contamination from the...

06 December 2018 762 3 View

How to assess the osmotic power of a substance?

Hello, I suspect that a molecule I am using is causing bacteria to experience osmotic shock. What chemical properties should I look for to compare this capability with those of other substances?...

06 August 2024 977 1 View

How to check pH of a high strength hydrogel?

We are working on biopolymeric hydrogels. Our system is highly viscous and sticky, and the gel formed are high in strength. We are unable to use pH electrode and pH strip. Please suggest an easy...

30 July 2024 942 2 View

EDTA buffer solution?

Hello everyone! How can i prepare EDTA buffer solution (pH 8.2) to determine GSH levels? Thank you in advance.

29 July 2024 4,374 1 View

Whether i can preserve leaves samples in 2.5% glutaraldehyde in 0.05 M phosphate buffer, pH 7 for coming several weeks for further LM study ?

I want to preserve the sample leaves for the LM study of the stomata and conidia analysis. Whether 2.5% glutaraldehyde in 0.05 M phosphate buffer with pH 7 can be applied for preservation.

29 July 2024 8,560 0 View

A positively charged and 10x his-tagged protein that doesn't bind to any chromatographic resin?

Hello everyone, I am currently working on a protein that is 10x his-tagged and positively charged (predicted pI=10.16). But when I tried to use Ni-NTA column to purify the protein, it's not...

24 July 2024 7,293 4 View

What is the principle/mechanism behind aging of carbon (graphite) containing refractory mix for isostatically pressed refractories?

The resin bonded carbon containing refractories are aged before use. How time of aging is determined? And mechanism behind aging.

23 July 2024 3,205 0 View

Prevent sedimentation and maintain the suspension of powder particles in the resin ?

Hello, I am experiencing an issue with mixing micro powder with resin in a 3D printer. The problem is that sedimentation occurs after a short period (about half an hour to an hour). Could you...

23 July 2024 1,672 0 View

What is the physical meaning of the magnetic scalar potential?

In cases where the rotational of the magnetic field H is zero, we can define this field as the gradient of a scalar function defined as the magnetic scalar potential (similar to the electric...

21 July 2024 9,633 4 View

Any relation between resin viscosity（mPa*s）and resin relative molecule weight？why the former is used in manufacture?

https://en.wikipedia.org/wiki/Viscosity

21 July 2024 3,089 4 View

Is it possible to conjugate an NHS ester to the N-terminus of a peptide ON RESIN?

So i have a few peptides (synthesized by SPPS and not cleaved from resin) and I want to add a maleimide label. We have NHS-PEG-Maleimide and from literature it is evident that the free amine group...

18 July 2024 4,075 3 View