ALP protocol in cell culture?

More Eduardo Pons-Fuster's questions See All

¿Cuáles son los errores mas comunes en la implementación de las políticas públicas?

Por ejemplo en el Perú, se ha trabajado una interesante política de seguridad ciudadana con claras directrices, sin embargo los resultados de sus indicadores son contraproducentes, además el gasto...

20 April 2024 9,405 2 View

Why are the future aspects of prefrontal functions neglected?

Friends, The difficulty to form and execute plans of action is diagnostic of large prefrontal lesions. The expectancy of reward activates many neurons, especially in orbital prefrontal...

24 November 2017 5,035 8 View

Measuring Nursing students leadership (SALI)

Have you used SALI (self assessment leadership instrument) scale? I have some doubts about the organisation of the items. Thank you all! Pilar

20 May 2017 1,158 5 View

Change in SAOS-2 morphology?

Hello, I´m currently doing cell culture using Saos-2 osteosarcoma cells . I have only split them 6-7 times, but they seem to have changed their morphology and they do not divide anymore. I thawed...

11 December 2016 8,799 3 View

Using CRISPR/Cas9 in NHDF?

Hi there! We have previously implemented a CRISPR/Cas9 design in Hek293 with great success (more than 60% cutting efficiency), but now we want to move the experiments to human adult fibroblasts...

25 July 2016 9,535 2 View

“The Executive of the Brain”: is this a useful concept?

The motor cortex is in the frontal lobe; the prefrontal cortex controls the main executive functions (executive attention, working memory, decision-making, inhibitory control); patients with...

13 June 2016 9,477 34 View

What is the minimal neural structure capable of storing a personal memory or item of knowledge in the cerebral cortex?

Memories and knowledge are acquired by mechanisms we don’t yet quite understand and mediated by limbic formations, notably the hippocampus (“old cortex”). After a period of consolidation, they...

22 May 2016 1,042 40 View

Why is working memory considered a special kind of memory instead of a state of memory?

Baddeley (1983) said it clearly, that working memory is the temporary retention of recent information for the performance of a task or the solution of a problem. Yet the future perspective...

17 April 2016 9,728 24 View

Can I clone GFP together with my protein without it being a fusion protein?

Hi! Could anybody tell me if it is possible to clone a GFP tag with my trasngene of interest without being a fusion protein, just a separated elements? Thank you.

11 February 2015 9,031 6 View

What method would you recommend to identify newly formed blood vessels in a tumoral formalin fixed paraffin embedded tissue?

We would like to quantify the newly formed vessels in slides of formalin fixed paraffin embedded tissues but the methods we are using now seem not to be useful enough. Any suggestions?

21 March 2013 6,957 25 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 2 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

Good Day, can I ask for your help on the questionnaire that I want to adopt?

I am currently looking for a questionnaire that relates with my study which is The Effectiveness of Organizational Culture towards employee retention rates

03 August 2024 3,505 6 View

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?

I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...

03 August 2024 3,851 1 View

s. Béatrice Marianne Ewalds-Kvist

Dear Eduardo,

see if this paper helps you:

Article Regulation of the human tissue-nonspecific alkaline phosphat...

Sherif Suleiman

Hi Eduardo,

Try the sigma alkaline phosphatase yellow pNPP liquid substrate system for elisa. Cat No. P7998-100ml. The protocol I do is simple. incubate SaOS2 with treatment and use RA as positive control. if using 96well plate, wash cells twice with PBS, add 50ul of the sigma solution directly on the cells, incubate at 37 in incubator for 45 minutes, read with spectrophotometer

Eduardo Pons-Fuster

Very useful! thanks!!