Rachel Lieberman

13 Questions 6 Answers 0 Followers

Questions related from Rachel Lieberman

Do murine isolated NK cells in culture need to be trypsinized for passage or are they floating?

I isolated NK cells from a mouse and they seem to stick to the cell culture plate. Should they be trypsinized for passage?

10 October 2016 6,127 4 View

How to detect growth changes by addition of conditioned media?

I want to do an experiment where I take conditioned media from one plate of cells and and add it to a plate of cells with a gene knockout and see if there are changes in phenotype. Can someone...

03 March 2016 5,266 1 View

How best to dilute camptothecin to i.p. inject 50 mg/kg into mouse?

I know camptothecin can be insoluble except in DMSO. Does anyone have experience diluting CPT in order to i.p. inject a high amount into a mouse?

03 March 2016 9,368 2 View

Can you use PE Annexin v apoptosis detection kit i with mouse lung tissue?

I have a PE Annexin V Apoptosis Detection kit from BD Pharmingen to detect incidence of apoptosis by flow cytometry. Can I use mouse lung tissue with this assay? If so, how would you suggest I...

02 February 2016 9,302 1 View

Can someone help me with genotyping PCR?

I am trying to genotype my transgenic mouse by following the protocol of a paper published on the original mouse data and cannot get a clear PCR gel. I want to genotype for the wildtype gene...

07 July 2015 2,009 12 View

Can someone help me calculate drug dilutions and dosing for i.p. injection?

I have an undiluted drug in powder form. I need to give it to my 20 gram mouse at a dose of 10 mg/Kg. The drug is docetaxel and its MW is 807.87922. The drug product sheet says that it is soluble...

07 July 2015 5,021 3 View

Does anyone have experience with an efficient assay for measuring sister chromatid exchange in cell culture? Any good commercial kits available?

I would appreciate an efficient protocol for measuring sister chromatid exchange or suggestions for a commercial kit available to purchase.

08 August 2014 7,665 3 View

What is the best method for transfection with RSV promoter?

What is the best method to transfect an overexpression plasmid with the RSV promoter? Should I make virus or directly transfect?

08 August 2014 9,527 2 View

Was I correct in using cDNA to check for mutant vs. wildtype p53?

I need to determine if my H226 cell line has wildtype or mutant p53. I extracted RNA, performed reverse transcription PCR and sent in 1 ug for sequencing to Retrogen . My sequencing failed and...

08 August 2014 895 16 View

How does one prepare 300 uM hydrogen peroxide from 3 wt % H2O2 solution ?

Confused trying to change weight % into molarity using sigma's hydrogen peroxide.

07 July 2014 5,058 4 View

Has anyone used a protease to chop up a large protein like BRCA2 in order to detect by WB?

Having trouble detecting BRCA2 protein by Western Blot. I used a 3-8% Tris-Acetate gel, 40 ug of protein and ran the gel for 4 hours. To transfer I ran the wet apparatus at 4C overnight at 25...

06 June 2014 522 2 View

Can anyone help with a Western Blot to detect levels of BRCA2 protein?

I have not been able to see any BRCA2 protein on my blots. I know that this is a high MW protein (390 kDa). Has anyone performed this successfully? Please let me know procedure specifics...

05 May 2014 4,676 5 View

Why am I seeing difference in cytotoxicity studies of wild type and gene knockout NK and CD8+ T cells without pre-activation in vivo?

Upon performing a cytotoxicity study using NK and CD8+ T cells isolated from wildtype and gene knockout Black 6 mouse splenocytes, the knockout immune cells induce more cell death and granzyme B...

01 January 1970 9,808 3 View