Can anyone help with my problem with periplasmic protein extraction by osmotic shock?

Giggsy Matinny @Giggsy-Matinny

08 November 2015 1 1K Report

I'm very confuse about time for incubation of protein after add sucrose buffer and osmotic shock buffer(MgSO4).

and volume of each buffer were added?

Buchs Natasha

Maybe you can find some help in this article

http://onlinelibrary.wiley.com/doi/10.1021/bp034030s/pdf

Good luck

Badges
Science topic

More Giggsy Matinny's questions See All

Question about plasmid digested by two-cut-site RE at MSC?

Here I have a basic question. My cutting vector run faster than uncut when cut with MscI and XhoI. Who can tell me it could happen in the normal situation?

12 February 2016 387 1 View

No title

What is the optimal IPTG concentration and incubation time after induction? Normally, My protein were induced with 0.5mM at 25'C for O/N but now I have limitation about shaker, so i'll to try new...

21 September 2015 9,707 3 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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 server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 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

Is there any one who can provide a donation of HPV detection and genotyping kits in exchange collaboration fors?

Warm greetings My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology (on Human papillomavirus). At this stage, I have almost completed a clinical sample Cervical swabs)...

25 February 2021 1,660 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

Is there any one who can provide a donation of HPV detection and genotyping kits in exchange collaboration forms?

Warm greetings! My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology focusing on Human papillomavirus). At this stage, I have almost completed a clinical sample (Cervical swabs)...

25 February 2021 3,612 3 View