Unable to detect a truncated protein variant, but I can observe a truncated cDNA from Sanger?

Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...

17 February 2021 1,130 6 View

Where can I find these two papers?

14 January 2021 1,572 3 View

After CRISPR and single-colony picking, cells reverted to WT?

Hi guys: I have been doing CRISPR knock-in and knock-out and one seemingly impossible scenario keeps happening. In the CRISPR knock-in attempt I tried to add a tag on a gene of interest, so I...

07 January 2021 190 3 View

Is there any ingredient I an add into formula to slow down gas generation from Vitamin C?

I know Vitamin C is unstable and tends to be oxidized quickly. The problem here is I made a Vitamin C beverage at pH 4-5, and it starts to generate gas in the packaging, especially at high...

13 December 2020 6,055 1 View

Contamination when culturing primary uterine endometrial cells?

Dear everyone I'm working on the endometrium of the uterus. I need to extract endometrial cells from the endometrial tissue. Usually, I will separate the tissue into 2,3 mm pieces and incubated in...

25 November 2020 7,428 3 View

What are the morphological changes of PC3 cells during autophagy and/or apoptosis?

I was performing wound healing assay seeding 300,000cells/well in 6 well plates with 2mL RPMI medium. At 0hr after intervention, the cells looked like the first picture. At 24hr after...

18 November 2020 9,671 2 View

Why use STOP solution in ELISA?

At the last step for data analysis, we always use a STOP solution to stop the TMB oxidized by HRP. Why do we have to stop the reaction? Couldn't we measure the color wavelength of...

17 November 2020 5,177 3 View

Why case-controlled study is more bias prone than cohort study?

I noticed from several articles saying that case-controlled study is more bias prone than cohort study (in the context of cancer early detection using molecular marker). I spent sometime to try to...

17 November 2020 1,878 4 View

Does PROCESS v3.5 by Andrew F. Hayes with SPSS Statistics 27 on MacOS 10.15.7 work?

After installing the PROCESS v3.5 tool, I ran a mediation model 4 with three variables (N = 125). The warnings below showed up on the output window, and the status at the bottom shows, "Running...

15 November 2020 3,930 14 View

How to perform statistic analysis for age range between two cohorts?

Hi, I was reading the paper (MicroRNA Expression, Survival, and Response to Interferon in... ). In Table 1, I'm wondering how did the authors calculate a p-value of 0.29 for Age? The age in Table...

20 October 2020 2,276 5 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

How does the gut microbiome affect the fibrotic process within the liver?

and Can the gut microbe be used for the treatment or diagnosis of liver fibrosis?

02 March 2021 557 2 View

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

Is there an appropriate CD8 antibody for IHC-P for staining human breast cancer tissues?

Hello, We are planning on staining Formalin-fixed paraffin breast cancer tissue sections (FFPE) with CD8 Ab via immunofluorescence (Tissue IF). Based on our existing protocol and imaging...

24 February 2021 9,578 4 View

Is it harmful to breed for Cre Homozygosity initially?

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...

24 February 2021 5,759 3 View

Is CXCR5 on T cells affected after a cell sorting?

I have tried to sort CD8+ CXCR5+ in the ARIA but the purity of my sample is about 50%. After sorting, I stain my samples to verify purity but it is low. So, I thought about "downregulation" or...

15 February 2021 8,034 7 View

How should I go about evaluating cytoplasmic and nuclear Arginase-1 activity?

Hello! I've stained some bright field Arginase-1 Hematoxylin slides, but I'm having a hard time finding any computed histology predicates for evaluating the intensity of the cytoplasm verses the...

03 February 2021 3,039 1 View

What's wrong with this fronzen section of mice melamona tissues?

I met a problem when I was doing frozen section for mice melanoma tissue. Based on the confocal results I got as attached, seems like the cell structures are destroyed (looks like a network) and...

31 January 2021 8,464 4 View

How do I avoid artifacts during paraformaldehyde fixation of mouse CD8+ T cells?

I work with mouse OT-I CD8+ T cells which have been activated in vitro with peptide-MHC and B7 proteins for 3 days. I've been trying to standardize protocols to fix these cells using...

28 January 2021 8,519 2 View