How can I overcome such problematic melt curve in qPCR?

31 December 2015 5,690 5 View

How to achieve better amplification in qPCR?

09 October 2015 6,293 9 View

How might I ensure RT-PCR consistency ?

06 July 2015 6,551 5 View

Why is my western blot black and without bands?

I am currently doing western blot for p-Akt. Below is my protocol: -- Blocking = RT, 2 hours with 5% BSA -- Primary antibody = p-Akt (1:2000), overnight, RT -- Secondary antibody = Anti rabbit...

04 May 2015 6,166 9 View

What is the difference between non-fat milk and BSA for primary antibody in western blot?

Some protocols mention primary antibodies should be diluted in 5%BSA whereas some protocols mention primary antibodies should be diluted in 5% non-fat milk. What is the difference? What is the...

04 May 2014 387 20 View

Is serum starve important for TGF-beta induced EMT and EGF induced EMT?

Many publications mention that cells were serum starved for 24 hours before adding TGF-beta or EGF to induce EMT. After serum starved, is it possible to treat cells with EMT-inducers in RPMI 5%...

04 May 2014 7,714 10 View

Do you know where I can find cancer genomic data on kyse50, kyse 110, TE2, and TE3?

I have found that cbioportal is a good website to study molecular expression level of certain proteins in certain cancer cell lines. Unfortunately, some cell lines are not in the database. Where...

03 April 2014 1,016 4 View

Can anyone help: my western blot result of a snail protein is higher than 30 KD?

The supplier COA stated that the snail protein is about 30KD. I ran the electrophoresis at 200V, 125mA for 45mins. I used 4-12% Tris Nu-PAge Gel. Why is the snail protein more than 30KD?

02 March 2014 8,085 7 View

How can I improve my western blot results for the optimal dilution for a Snail-1 antibody?

I am currently comparing snail-1 expression in esophageal cancer cell lines. My protein load was 30-40 ug, the primary snail-1 antibody (cell signaling) was 1:1000, and the secondary anti-rabbit...

02 March 2014 8,669 17 View

Which scale is better for the Measurement of Fluorescent Intensity?

Which scale is better, Linear scale or log scale?

02 March 2014 7,258 7 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 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

No title

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

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

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

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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