Adding more media to bring down O.D before protein induction?

Hello folks, I found that my E.coli BL21 DE3 star cultures grew to O.D. 1.2 before I could induce with 0.5 mM IPTG. I normally induce at O.D. between 0.4 to 0.8 when they are in exponential...

07 August 2018 5,750 0 View

White cloudy precipitate during Ni-NTA purification?

Hello folks, I had a N terminal His-GST-TEV cleavage site tagged protein (15mg) on dialysis over ~40 hours in the presence of TEV protease (1mg) with a buffer change in between. Buffer is 50 mM...

06 July 2018 1,277 5 View

How to identify degraded RNA and non-denatured RNA products on a PAGE-UREA gel?

08 September 2016 1,647 0 View

Unusual Chloramphenicol absorption in sucrose gradient on Isco fractionator?

Hello there, I have prepared 10-55% sucrose gradients containing 1mM Chloramphenicol and run them on an Isco sucrose gradient fractionation system, but the absorption is too high to record on the...

03 April 2016 9,871 3 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

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03 March 2021 5,360 2 View

Ester cleavage conditions?

Hi all, I need to cleave the ester in Ethyl 2-amino-1H-imidazole-5-carboxylate to make the carboxylic acid and retrieve the product from solution to use in downstream applications of linking to...

01 March 2021 8,766 3 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

How can we raise the figure when we writing a paper with Latex ?

Hi, everyone. I'm a starter to write a paper by Latex. But I have some difficulty using it. I'm writing the paper with 2 columns. And, there is one figure that occupies the 2 both-sides. I want to...

26 February 2021 3,738 3 View

What is the best way to calculate the angles of twist for concrete columns subjected to torsional moment using finite element software?

In Finite element software like ANSYS , there is difficulty in obtaining direct results of twisting of concrete members because SOLID65 which is the unique element designed to represent concrete...

26 February 2021 3,806 2 View

Compute for the total volume of concrete needed for the concrete pouring of the foundation including its column up to NGL only as shown in the plan?

Looking for the quantity of cement, sand, and gravel of footings and columns. Thank you!

25 February 2021 3,092 2 View

No amplification data for qPCR on MxPro?

I have set up a qPCR run on MxPro with SYBR green and ROX as a reference, however, when I go to the analysis section the program seems to only collect data for the dissociation curves and not the...

25 February 2021 8,039 1 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

Can I use TCA instead of TFA for protein analyis in RP-HPLC?

I am trying to analyze lipase using the reverse phase-HPLC C18 column. In the literature, 0.1 % TFA was and 60% ACN was used. Can I use TCA in place of TFA? If any other methods are available to...

25 February 2021 2,388 3 View