How to troubleshoot the IF staining problems in skin tissues?

Hi We are working in the area of psoriatic diseases. We are everytime getting the problem in expression of protein in dermis. We unable to see the expression in epidermis. Even, we did IHC too...

10 November 2018 9,096 1 View

How to process the mouse skin tissues for histopathological evaluation?

Hi everybody, In our studies, we have used BALB/c mice for antipsoriatic activity evaluation. In that, we unable to get the proper tissue sections for histophathological and immunofluorescence...

05 June 2018 798 3 View

What are the biological assays for drug and TNF Alfa binding studies?

Dear sir/madam, Please suggest the biological assays which need to prove the drug and TNF Alfa binding studies. Thanks.

05 June 2018 5,334 1 View

How to disperse ZrO2 nanopowder in vehicle suitable for its use in cell culture.....

I'm trying to disperse it in Distilled water, and used sonication and homogenization approach to reduce its size but still getting PDI more than 0.5 and size more than 600nm

07 August 2017 5,069 4 View

How can I conjugate the miRNA with cy5 dye for tracking the transfection?

Hi everybody, I would like to transfect the anti-miR to the cancer cells. I have purchased it from sigma. I have the lipofectamine 2000 as a transfection agent.But I don't know the procedure to...

05 June 2016 1,110 2 View

How can I give the anti-miRNA to the athymic nude mice?

Hi everybody, I want to give the anti-miR as a therapeutic agent for the tumor bearing mouse. I have purchased the anti-miR from sigma Aldrich. Is there any standard protocol to deliver the...

05 June 2016 8,527 2 View

For getting Post-Doc in top universities, is there any requirement of guide profile and institution name?

10 November 2015 1,007 2 View

What are the requirements to maintain the cancer stem cells?

We have normal in vitro facilities to culture the normal cancer cells. I want to work on cancer stem cells. Could you please suggest me what are the effective methods to maintain the cancer stem...

09 October 2015 5,371 5 View

Could you please tell me the methods to isolate cancer stem cells without the application of FACS?

I want to isolate the cancer stem cells from cancer cell lines. But most of the people are isolating the cancer stem cells by using FACS. But in my lab it is not available. So I want to use...

09 October 2015 8,478 5 View

Why do I amplification in RT-PCR in RNA-free water?

I am using Taqman miRNA assay kit for miRNA amplification. But I am getting the peak for water control also. I have repeated by using new purchased RNAse free water, even-though I am getting same...

04 May 2015 5,874 7 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View