Jifan Yuan

9 Questions 7 Answers 0 Followers

Questions related from Jifan Yuan

How to improve my BrdU immunostaining?

Hi everyone, I am working on BrdU labeling and immunostaining recently. I have tried HCl hydrolysis, it didn’t work so I used 60 degree overnight incubation in citric acid ph6 to denature DNA,...

25 January 2021 650 3 View

Any tips on Fab blocking of mouse tissue?

Hi, everyone, I am working on immunostaining of bone marrow and growth plates of mice. And I have to use an mouse monoclonal Ab to do this staining. However, with only secondary goat anti-mouse...

22 January 2021 10,031 3 View

Any suggestions on 50um cryosection immunostaining (growth plate) ?

Hi everyone I am working on immunofluorescence of cryosection of murine growth plates. The Ab worked well on 8um cryosection. But when I stained 50um section, I found the fluorescent signals...

04 January 2021 9,735 3 View

How to reduce the false positives of GST pull-down?

Hi everyone, recently, I am going to study two protein interaction by GST pull-down. I have protein GST-A and MBP-his-B expressed by BL21 through IPTG induction. Initially, I found GST-A can pull...

08 April 2019 2,999 0 View

Questions about IHC and tissue fixation, embedding method?

Hi everyone, just ask some questions that I can't understand. It is a common sense that fixation block antigen, especially for the tissue that should be fixed for a long time (such as thicked skin...

10 October 2018 3,354 6 View

Co-immunoprecipitation, protein a/g contamination?

Hi I am recently doing Co-immunoprecipitation experiment. And I can always find two bands at 35kD and 40kD (I use rabbit polyclonal antibody as primary antibody of WB) which are strong enough and...

09 April 2018 4,257 3 View

Pfa in skin cryosection immunofluorescence?

I have a lot of questions about murine skin tissue immunofluorescence (IF). 1. What causes autofluorescence in skin tissue? Even though I don’t use PFA to fix before embedding Into OCT,...

22 November 2017 2,857 1 View

Is there anybody familiar with cre-ert-gfp mice?

I have been working on this for over one year. After I getting a lot of target cre-ert-gfp mice, I try to localize a group of cells expressing cre-ert-gfp by either immunofluorescence using GFP...

14 October 2017 1,145 2 View

Background staining in methanol and acetone-fixed cells

I am doing IF on methanol-fixed cells (The antibody does not recognize the antigen if I use PFA to fix cells). I get high 'perinuclear' background even in control (only fluorescent secondary...

01 January 1970 5,960 3 View