Debosree Pal

10 Questions 21 Answers 0 Followers

Questions related from Debosree Pal

My antibody is not pulling down the protein of interest during immunoprecipitation. What should I do?

I have an in-house raised antibody against my protein of interest which I have purified from the serum by the caprylic acid method and further confirmed on a sds gel. I wanted to use it for...

12 December 2015 2,717 7 View

Has anyone worked on embryonic germ cells as a model system?

I want to work on embryonic germ cells. Can anyone suggest from where I can procure these cells? Also, does anyone know the difference between embryonic germ cells and primordial germ cells

02 February 2015 1,380 1 View

Can R1 embryonic stem cells be grown in 2i+LIF medium?

Hello, I am growing R1-ESCs in high glucose DMEM, 15%FBS, sodium pyruvate, non-essential amino acids and L-glutamine. Can these cells be grown in 2i+LIF medium? How many passages do I need to grow...

10 October 2014 7,223 4 View

Can anyone share with me a protocol for the differentiation of mouse embryonic stem cells into primordial germ cells?

Hi, I am looking for a protocol to differentiate mouse embryonic stem cells into primordial germ cells. Can anyone provide me with a convenient protocol? How long can the differentiated cells be...

10 October 2014 9,698 5 View

Does anyone have experience with an inducible lentivirus based shRNA expression system for mouse embryonic stem cells?

I am planning to perform shRNA -mediated knockdown of my gene of interest using an inducible lentiviral based system. I am working on mouse embryonic stem cells. Can someone suggest a suitable...

08 August 2014 6,017 7 View

Can a tet-off sysytem be replaced by a tet-on one?

I have a plasmid in which the TetO2 operator sequence is present in the promoter for the gene. The manual says that it is supposed to be used for the tet-off system. Is the binding sequence for...

07 July 2014 8,346 1 View

Can anyone suggest a suitable tet-on inducible lentiviral plasmid for shRNA expression?

I want to express shRNA against my gene of interest in a tetracycline regulatable manner (tet-on system). Can anyone suggest some suitable plasmids for the same? I have shRNAs with 5'BglII and...

07 July 2014 3,533 3 View

Can anyone suggest how to store hybridization buffer?

I am using a hybridization buffer for in-situ hybridization which has the following components: 50% formamide, 5X SSC, yeast RNA, heparin and 1% SDS. Storing the buffer at 4 degrees or -20 degrees...

05 May 2014 3,412 6 View

Does the method of homogenization of tissues bring about a change in downstream PCR results?

I was trying to see the expression of a gene in post-implantation mouse embryos from 9.5-13.5dpc. The preferred technique is obviously whole embryo in-situ hybridization but, I first thought of...

05 May 2014 2,236 4 View

How tricky is it to isolate and culture primary mouse embryonic fibroblasts?

I have been trying to culture mouse embryonic fibroblasts for quite a while now but have been facing a lot of problems with the same. I isolated the fibroblasts from E13.5 old mouse embryos (CD-1...

04 April 2014 7,683 7 View