Why is there expression in shaker but not in fermenter?

I have a protein of interest that is expressing in Pichia. It is expressing well in the shaker flask, but when I apply the same expression methodology but now in a bioreactor I get minimal...

03 April 2018 6,866 8 View

Why is my protein precipitating at lower ammium sulfate cuts NOT at higher?

Procedure: Split supernatent into three individual samples A, B, C and do 20% NH4SO4 cut on sample A, 40% cut on sample B, and 60% cut on sample C. Add granulated ammonium sulfate NOT a saturated...

11 December 2017 8,061 8 View

Which insertion should I use in Pichia pastoris?

According to Invitrogen pichia pastoris cloning system using pPIC vectors there is an option to insert into AOX1 or HIS4 when using GS115 or KM71 strains. In both cases the SAME phenotype is...

09 October 2017 5,128 2 View

Using CMFDA vs DCFDA for ROS Detection?

Structurally they are similar and my understanding is that the OH groups are oxidized which ultimately modulates fluorescence. Both of these molecules are identical in that respect so why is DCFDA...

04 May 2016 467 2 View

I am looking for a protocol to knockout a gene in ovarian cancer cell line.

The old school way (plasmid/homologous recombination) not using CRISPR. I can't find anything relevant right now. Does anyone have a PDF or some manuscript describing a step by step sequence? It...

03 April 2016 4,724 1 View

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02 March 2021 3,060 3 View

Where can I find life cycle inventory or assessment of non woven materials?

Dear network, I am looking for the LCA or LCI or at least the energy balances of the non woven fabrics process for my master project, in specific for natural fibers, yet I have found...

02 March 2021 9,662 1 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

01 March 2021 8,087 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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