What is the best way to fluorescently label TLR3?

Dear all,

I want to build the Sorafenib Tosylate resistant hepatocarcinoma cell lines, HepG-2 and Hep-3B, does someone have experience about this?

I have no experience about this, so I want to get some suggestions from you.

How to start? and how to continue?

Thanks.

Hi All,

I have SK-N-SH cells I would like to differentiate, what protocol do you normally use? I can see that people do it in different ways, I want to know which method gives you the most consistent and robust differentiation efficiency. Thank you!

I need to insert RFP or GFP gene into E. coli genome to have permanent fluorescent cells.

Is there any helpful kits and recommended chromosome position for this purpose?

I am not intending to use anything to make the sample fluorescent or should I use Calcium Hydroxide+Phosphoric Acid, and then doping with Fluoride salt.

Any idea? 

Does anybody know how the process should be done in detail? I am a little bit confused with concentration of each component....

I would like to enhance the fluorescent signal of mRuby3 in brain slices. I have only found antibodies good for WB. Did anyone try them on tissue? Is there a good antibody for IHC?

Dear molecular biologist experts,

I am combining three probes that should be revealed in a multiplex real time pcr with the system LC480. Do you have any experience with dramatic drop down of fluorescence in a triplex with the following probes and channel?

Channel:Exc-Det

FAM : 465-510

LC610 : 533-610

CY5 : 618-660

protocol:

(95°C 5 min) x 1 cycle

(95°C x 10 sec - 60°C x 45 sec - 72°C x 1 sec) x 40 cycles

(40°C 10 sec)

Let me know what do you think about

I am trying to purify a N-terminally GST tagged protein which is very well expressed in E.coli membrane. I was successful in extracting it out from the membrane using sarcosyl/detergents but the protein is somehow having low binding onto the column, most of the part seen in flow through. However I was able to purify soluble GST with the same buffers which I used for the protein of interest. What could be the reason for protein low binding onto column?

THP1 cell line is giving negative result for CD 58 expression in FACS analysis.

I am trying to measure the fluorescence of cells at plasma membrane using ImageJ. Could anyone help me how I could do it please. I attached a sample image also.