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...

11 December 2015 2,638 7 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...

09 October 2014 9,618 5 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...

09 October 2014 7,158 4 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...

07 August 2014 5,959 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...

06 July 2014 8,272 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...

06 July 2014 3,461 3 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...

04 May 2014 2,166 4 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...

04 May 2014 3,316 6 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...

03 April 2014 7,618 7 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

After a reference from Int J Body Compos Res, can any body help?

Hi, I am after the reference below, my library says it cannot obtain a copy either locally or internationally, any help appreciated! Chris Wang ZM, Heshka S, Wielopolski L, Pi-Sunyer FX, Pierson...

03 March 2021 6,193 1 View

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

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01 March 2021 3,905 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

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

01 March 2021 8,087 3 View