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Questions related from Joselyn Allen
My lab is venturing into new territory by testing the affects of different antibodies on bacterial uptake/phagocytosis by macrophages. Does any one have a general phagocytosis protocol, especially...
03 December 2022 7,791 3 View
Is it mandatory that ELISPOT plates be pretreated with Ethanol? Has anyone performEd an ELISPOT without doing this step and still got good result?
03 October 2022 3,286 4 View
Hello, I would like to use a mouse cell line to study the effects of the gluconeogenesis pathway. AML12 has been mentioned to not be the best model but others have stated otherwise. I would like...
07 November 2021 8,425 0 View
I am working on a project which involves evaluating bile acid profile in mice. We have repeated the experiment and have noticed high variability between mice within a given group and the trends no...
27 August 2021 678 0 View
I would like to evaluate the targeting efficiency of my crispr/cas9 model in mice and would like measure the aav8 genomic copy number in different tissues including liver, heart, spleen, etc? What...
29 July 2021 944 6 View
Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...
25 February 2021 6,214 3 View
I am optimizing a CRISPR/CAS9 system in vitro before using it in vivo. I would like to transfect AML12 cells with my pAAV-sgRNA-vector to validate editing efficiency before generating virus.
27 January 2021 9,087 2 View
I recently introduced a Lenti-Puro Guide vector containing a specific sgRNA sequence into a cell line. I would like to sequence the ddCas9-sgRNA cell lines to confirm that each sgRNA is accurate.
05 February 2019 6,933 3 View
I am performing a kill curve with puromycin, ranging from 0.5 ug/ml to 10 ug/ml. It has been 6 days from initial treatment and 100% of the cells have not been killed as yet. Cells have died but...
12 September 2018 9,309 0 View
I recently analyzed RT PCR data for NASH livers. Oil red O staining revealed that my experimental group has exacerbated NASH/ lipid content. This group also demonstrates lower RNA expression of...
20 May 2017 3,980 2 View
The adipose have been sectioned and stained with H and E. Of couse F4/80 is required, however I would to crudely analyze these sections.
16 February 2016 7,149 1 View
LPS is shown to induce SREBP expression in the literature, however, there is inconsistency in the literature methods that states to perform the experiments in serum free media and other papers...
12 November 2015 9,488 3 View
I am stimulating primary hepatocytes with TNFa and evaluating the expression of key genes; however I have been using recombinant human TNFa on mouse hepatocytes. I am not sure whether I should...
13 August 2015 6,455 1 View
I am using stimulated primary hepatocytes to detect alterations in AMPK activity. However, a paper published that serum starvation can increase AMPK activity in different cell types. Although,...
07 August 2015 3,616 3 View
I would like to assess the editing efficiency of designed guide RNAs by transfection prior to establishing a stable KO cell line (via lentiviral infection). HEK293T cells are very efficient when...
01 January 1970 8,179 2 View
Does anyone have a strategy for calculating ELISA titers from OD dilutions (without the use of standards)? The current method of my lab is through graphpad analysis (Sigmoidal 4PL, X). Using the...
01 January 1970 5,570 0 View
