Is anyone working with Fabry cell lines?

I am currently looking for laboratories or cell line repositories that are working or have worked with Fabry disease who are willing to provide me some transformed Fabry cell lines preferably...

07 August 2017 2,191 2 View

How can I induce tumorigenesis via in situ injection?

I would like to induce colorectal cancer via injection of cancer cell lines into mice models. So far I've found literature regarding the number of cells to be injected, but I'm not sure which part...

03 April 2015 4,444 6 View

Should I PCR the whole gene or not?

I am going to manipulate KRAS gene from mammalian cancer cell lines for vaccine production, and the gene codes for the KRAS protein (189 amino acids). I've already chosen my own antigenic region...

10 November 2014 8,611 4 View

How many types of polyphenol oxidases (PPOs)?

I'm writing my literature review on PPOs now, and facing a problem of identifying the type of PPOs. I found out that there are tyrosinase, catechol oxidase, and laccase, being the 3 major...

11 December 2013 962 3 View

Any protocols in enzyme purification that retain high enzyme activity?

I'm going to work on polyphenol oxidase (PPO) extracted from a fruit peel, and from what I read recently I came out with a series of purification steps involving ammonium sulphate fractionation,...

04 May 2013 1,145 22 View

How can I concentrate a pooled fraction that contains my desired enzymes?

For example after running a column chromatography I realized that five of the collected fractions contained high enzyme activity (after running an assay on each). So I've pooled the five fractions...

04 May 2013 4,557 8 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 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

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View

How to reduce static electricity of powder after lyophilization?

I am working on synthesizing and then purifying peptides using the SPPS system. After purification of the peptide, the peptide powder is obtained using a freeze dryer. When measuring the powder,...

22 February 2021 10,099 5 View

How to calculate the fraction metabolized of a drug by an enzyme using clearance(ml/min) values of the metabolite formed?

I am trying to find the fraction metabolized of a drug by CYP and UGT enzymes. I am unable to get the values of fraction metabolized (Fm) directly from the literature. Can we calculate the Fm of a...

21 February 2021 5,379 2 View

Does anyone have a working Guanidine Hydrochloride purification protocol for synthesized ssRNA?

I am trying to in vitro transcribe some fluorescein labeled ssRNA. And I want to purify the ssRNA from the free NTPs and the enzymes (RNA polymerase and DNase I). Due to the characteristic of the...

18 February 2021 4,392 4 View

Purification of proteins from baculovirus-infected insect cells?

18 February 2021 6,665 6 View