Why Are Liver Cell lines Not Sensitive to Cyclophosphamide?

Cyclophosphamide is supposed to be activated by Cytochrome P450. I've treated Huh7 and PLC liver cancer cell lines with Cytochrome P450, but there is no sensitivity whatsoever, even at 500uM....

31 December 2016 931 6 View

PH versus pD versus pT of phosphate buffer?

07 August 2016 2,129 2 View

PKa of carbanions with Protons, deuteron and tritions?

I have an Emmons-Horner-like agent (triethylphosphonoacetate) that I would like to label with tritium. The alpha proton is readily exchangable in NaHCO3/D2O; I presume the same would work with...

07 August 2016 1,162 3 View

Can an R-enantiomer specific enzyme be engineered for the S-enantiomer?

Many enzymes have preference for L versus D or R versus entiomers. Is there a precendent in the literature for changing, engineering by amino acid alteration, a D-selective into an L-selective enzyme?

02 March 2014 1,550 3 View

Does anyone know of a mouse specific antibody for a housekeeping gene?

I have xenografted human tumor lysates that I am running western blots on. I need to determine the extent of mouse cell contamination in these lysates (I don't have slides for the tumors). I am...

05 June 2013 228 8 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

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

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 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

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 2 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 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