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Questions related to Protein Expression
Does anyone have a suggestion for an expression and purification protocol? I’m going to use BL21(DE3) E. coli strain and a pET-15b vector. All protocols seem to be very general and I don’t want to...
05 May 2018 5,317 2 View
Hi everyone, I am looking for mammalian expression system that allows for titration dependent expression of proteins. Meaning: a system that allows me to control the level of protein expression...
17 April 2018 6,557 4 View
Hello, I am looking at protein expression across mouse retina. We use littermate controls so many times our sample size is limited to 4-8 samples. To ensure protein does not degrade, we usually...
04 April 2018 4,296 2 View
04 April 2018 2,683 5 View
In pET sytem manual, it is stated that after protein expression and during harvesting cells of BL21 (DE3), you should wash the pellet with 20mM Tris-HCl pH 8. Does anyone know the reason for this?
04 April 2018 5,335 4 View
Can i use gfp or egfp antibody to detect copgfp protein expression when i do immunofluorescence?
04 April 2018 460 3 View
HI, I am working to express proteins from pichia. Can I flash freeze the cells and store at -80 degree? Does protein get denatured after freeze and thaw? Do I need to proceed to protein extraction...
04 April 2018 6,824 6 View
I want to collect endometrial tumor samples and subcutaneous fat tissues from patients for mRNA and protein expression, DNA methylatio analyses. I am really new and this is a very preliminary...
04 April 2018 1,040 3 View
04 April 2018 4,424 3 View
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...
02 April 2018 6,254 7 View
Hi everyone, I know that TLR signal can recruit the myd88 to cell membrance from cytoplasm. Here I observed thatTLR4 protein slightly increased while total myd88 protein expression decreased. Do...
28 March 2018 2,214 2 View
Is immunofluorescence staining or western blot is more sensitive to study the protein expression?
26 March 2018 6,049 8 View
Hi, I am planning to perform small scale expression in E.coli system, and I would like to use 12- or 24-well sterile plates (deep-well type). Are you aware of something like that available on...
19 March 2018 5,188 3 View
Dear All, I have been doing the expression and Purification of GST-HSP90M domain in BL21 9DE30 strain. I used to use 2x YT broth, 16 degree C induction with 0.2 mM IPTG for overnight induction....
19 March 2018 1,931 0 View
I have cloned a gene for a 10 kDa bacterial protein in pET28a (to be expressed as a C terminal His tagged protein) and transformed into DH5a. I have confirmed the clone by restriction digestion...
06 March 2018 6,738 4 View
I want to test different strains to increase protein production in yeast and want to limit it to s. cerevisiae
03 March 2018 1,133 4 View
03 March 2018 6,679 4 View
I cloned a gene(esat6) in pet25b vector. But after IPTG induction there is no protein expression. Why? I confirmed the clone by sequencing
03 March 2018 10,041 5 View
03 March 2018 5,067 6 View
e.g In a sample of breast cancers. tumour grade and Lymph node status (positive/negative) tumour grade and size, , average cells expressing a certain protein receptor status (positive/negative)...
03 March 2018 1,210 1 View
I am trying to optimize the expression conditions for S. cerevisiae yeast cells, I need high expression for downstream screening, and was wondering if anyone of you has tried higher Galactose...
03 March 2018 1,499 3 View
03 March 2018 8,787 2 View
I have been expressing an intrinsically disordered protein (IDP) and have been having issues purifying it. The protein is overexpressed in T7 promoter E.coli cells. The protein has a 6His tag and...
20 February 2018 3,650 6 View
I have a clone of my protein coding ORF in pGEX-4T-1 vector which should produce a fusion protein of ~115kDa. Snapgene Analysis of the clone (sequence) shows that the clone is in proper...
05 February 2018 2,415 6 View