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

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Markers of mitochondrial activity in FFPE tissues?

Hello everybody, Does someone have any experience with assessing/extimating mitochondrial activity on FFPE tissues? Precisely, no TOM20, ATP5B, etc: we are trying to get an approximative idea of...

24 February 2021 4,618 4 View

Can a Golgi-stained dendritic spines picture processed by Image J be directly used for publication?

24 February 2021 3,348 2 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