Cancer Research

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Questions related to Cancer Research

How to preserve a fresh tissue after surgery for future molecular analysis (cancer research)?

After surgery, what is the best condition to preserve fresh tissue samples from cancer patients for future molecular analysis? For instance: glycerol, formol, saline buffer? We will analyze these...

27 October 2016 7,048 4 View

Angiogenesis assay using matrigel

I am using growth factor reduced basement membrane extract ( 10mg/ml Matrigel from ATCC) to compare 2 conditioned media (CM) from HUVEC transfected with a siRNA control negative or siRNA against...

26 October 2016 1,245 3 View

Hi-- I'd like to extract CTLs from primary tumor and test proliferation profiles. any experience with harvesting viable t cells from primary tumors?

Harvesting viable immune cells from primary tumors in mouse model

26 October 2016 5,102 3 View

Can anyone spot any possible problems with this approach to diagnosing colorectal caner?

Take a sample from the suspected area, polyp, ulcer etc. Extract RNA from these cells. Convert to cDNA and do real time PCR to determine whether the oncogene VAV3 is over expressed? (VAV3 is...

26 October 2016 9,713 3 View

What lysis buffer is recommended for use with Benzonase when nuclear lysis is desired?

Hello, I would like to use Benzonase in my lysis buffer to reduce viscosity. I have been using RIPA buffer (50mM Tris, 150mM NaCl, 0.5% Na deoxycholate, 1% NP-40, 0.1% SDS, pH 8.0) to lyse my...

24 October 2016 2,936 3 View

Hello, can anyone assist me about the exact conditions to develop a western blot, using Luminata forte?

I just need an assistance about how to develop a blot , using luminata forte and Xray cassete? Like the conditions in which the developer, fixer or the luminata forte to be opened... Is complete...

23 October 2016 3,696 3 View

Does anyone know what is the Edu incubation dose for ID8 cells when trying to do Edu/propidium iodide assay for cell cycle/viability assessment?

Hi, I want to do a cycle/viability assay for ID8 ovarian cancer cells after treatment with the epigenetic drug Decitabine, on various doses. Has anyone done it and if yes, do you know the optimal...

20 October 2016 4,399 3 View

Polysome profiling centrifugation parameters to separate polysomes better?

Hi, I have a question regarding the parameters (speed and length) during sucrose gradient centrifugation for polysome profiling. I'm attaching one typical chromatogram from my experiments. The...

19 October 2016 2,418 2 View

What experiment can I perform to know how much time can my PYVMTs be without FBS and still be 99% viable?

What experiment can I perform to know how much time can my PYVMTs be without FBS and still be 99% viable? I am currently working with PYVMTs and I need to know how much time can my cell line be...

19 October 2016 4,253 3 View

List of Breast Cancer associated proteins?

I wish to carry out an exhaustive study of all the proteins which are involved/associated in/with breast cancer (associated directly or indirectly both, with regards to its progression, increase...

16 October 2016 1,835 4 View

Focus position not found while using olympus fluoview microscope. Has anyone faced the same issue?

Hello everyone, I have just started using Olympus fluoview for confocal microscopy and I don't know much about microscopy. I plate my cells (HeLa) in 4 welled Chambers slide. Stain the cells with...

14 October 2016 5,323 5 View

MAPK signaling pathway in cisplatin induced apoptosis Jurkat T-cells?

Dear fellow researchers, I am working on a project to globally determine the phosphorylation quantitative profiles of cisplatin-induced apoptosis in Jurkat T-cells. I have 4 biological replicates...

14 October 2016 2,471 3 View

Does Qiazol enables the extraction of proteins and DNA from the same sample along with RNA extraction?

I want to extract proteins and RNA from tissue samples. TRIzol reagent is not available in our labs. We have Qiazol instead? Does it do the same job? Are these products comparable in efficiency?...

14 October 2016 815 3 View

Quantifying exosomes for western blots?

Has anyone experienced difficulty in quantifying exosomes to be used for western blots? Ive been collecting exosomes from conditioned media and purified them using ultracentrifugation method. I...

13 October 2016 6,058 19 View

Labelling monoamines with dansyl chloride?

Hi, I'm trying to observe where dopamine localises at the cell surface of my cells of interest but fluorescent dopamine is very expensive to buy. I've read online that I should be able to label...

13 October 2016 5,597 2 View

Death in human primary CD8+ T-cell culture: advice?

Hi everyone, I am working on human Primary CD8+ T-cells and on their behaviour under various conditions. once activated in a OKT-3 coated plate and anti-CD28 for one hour, i put my cells in...

12 October 2016 6,637 3 View

Are 0.45 micrometer PTFE and Nylon syringe filters autoclavable?

Does autoclaving changes the pore size of these membrane filters

10 October 2016 6,351 2 View

Issue with protein overexpression?

Hi, I am using a lentiviral vector to over express a protein in AML cells. I have cloned the flag-gene into the EIFalfa-MCS-SV40 vector. When I use this vector to simply transfect 293 cells, I can...

07 October 2016 1,095 3 View

Could someone help me to choose the right concentration and time of neutralizing IL1beta antibody after LPS treatment to block its signalling?

Thanks!

07 October 2016 4,825 3 View

Why negative control plates got a lot of colonies using Single-step digestion-ligation cloning of CRISPR plasmid?

Based on Fengzhang lab protocol provided on their lab website,I performed the one step digestion and ligation, I got a lot of colonies on negative control plate. I am wondering the cut sequence...

05 October 2016 10,040 1 View

Best way to measure CK2 acitvity ?

Looking for best way to measure CK2 activity in cardiomyocytes. Western hasnt been working great want to use a different method. But it cant be radiometric assay. Thanks!

05 October 2016 3,276 2 View

Western Blot - How much dry milk in the antibody solution?

Hey guys, i'm looking for a general answer. My actual lab is blocking their WB with 5% Milk in TBST and then using 0,5% for the 1. and 2. antibody solution. My last lab was using 5% milk for...

05 October 2016 6,718 12 View

How could we understand the synergsitic Tumorigenesis effect?

We usually see that cancer progression is increased till death of a patient. Sometimes, this progression becomes fast after chemotherapy. As a result, patient dies due to synergistic effect of the...

03 October 2016 9,459 3 View

Should ribosomal RNA depletion for RNaseq (ribominus kit) look like that?

Hi I am depleting the ribosomal RNA of my samples with ribominus kit for ion proton libraries. After running a pico chip in the bioanalyzer I see a clear depletion of the main 18s and 28s (just...

30 September 2016 8,910 1 View

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