Protocol for Inducing Necroptosis in Cell Culture ?

Hi I am looking for a protocol (with appropriate dilutions, concentrations, quantities) of inducing necroptosis in in vitro cell culture. I have found protocols using TNF alpha dosing and TAK1...

31 December 2019 5,798 3 View

How can one model/code for Momentum Capture in ABAQUS to capture waveforms at particular time points?

I am trying to model a waveform through different bars in Abaqus to mimic a SPHB. I wanted to add momentum capture so I can get values of the waveform at different time points. How does one do This?

03 April 2019 8,554 2 View

Has anyone got an experience in using Bovine Thrombin from sigma?

- particularly this one http://www.sigmaaldrich.com/catalog/product/sigma/srp6556?lang=en®ion=US. Can it be used for cell culture? Does it need purifying before use?

04 May 2016 1,752 2 View

Problems with Bovine Fibrin Clots?

Hi I am using Fibrinogen (10mg/ml) and Thrombin (1000U/ml) to form fibrin clots. I found that after getting good cross linking initially the clots dissolve to liquid if left in incubator...

04 May 2016 7,107 5 View

Anyone have a good methodology for Agar Gel for adding on top of a cell culture dish (and that allows you to pour/pipette as a thin layer)?

01 February 2016 9,292 1 View

For Fibrin Matrices is >2000 NIH/ml best - or can one use 600-2000NIH/ml in order to produce 1000U/ml concentration (Thrombin from Bovine Plasma)?

- used for making matrices to seed neonatal fibroblasts.

01 February 2016 4,145 1 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 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

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Why the nucleic acid gel melted after running for 30min?

Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...

22 February 2021 9,896 3 View

Sanger Sequencing results not matching experimental results of Restriction Digestion, colony PCR and protein expression?

I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...

18 February 2021 6,736 4 View