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Questions related to Protein Expression
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...
04 April 2018 2,652 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 5,310 4 View
Can i use gfp or egfp antibody to detect copgfp protein expression when i do immunofluorescence?
04 April 2018 435 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 6,784 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 1,018 3 View
04 April 2018 4,408 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 6,230 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 2,191 2 View
Is immunofluorescence staining or western blot is more sensitive to study the protein expression?
26 March 2018 6,029 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 5,173 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 1,904 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 6,709 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 1,100 4 View
03 March 2018 6,656 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 10,010 5 View
03 March 2018 5,037 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 1,179 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 1,472 3 View
03 March 2018 8,750 2 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 2,396 6 View
I use the bac to bac expression system to express my target protein whic is secreted to supernatant.But the expression level is too low,who can give me some suggestions to impreove the expression...
02 February 2018 4,601 0 View
Need suggestion about this phenomena. If intron gets retain in processed mRNA. What would be the possible out comes in protein expression? Thanks in advance
02 February 2018 4,193 2 View
02 February 2018 4,633 7 View
Hello. I'm planning to perform the plasmid transformation efficiency using Cas9 protein in bacterial strain which has Type I-F and I-E CRISPR-Cas system. I transformed the pCas9 coding the...
01 January 2018 648 0 View