César Enrique Rivera —BIO-PROTOCOL

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Angela De Sisto Institute for Advanced Studies Foundation, IDEA, Venezuela
1 protocol

Ysvic Inojosa Institute for Advanced Studies Foundation, IDEA, Venezuela
1 protocol

Yusibeska Ramos Institute for Advanced Studies Foundation, IDEA, Venezuela
1 protocol

Laynet Puentes Energy and Enviroment, IDEA, Venezuela
1 protocol

César Enrique Rivera

Graduate, Energy and Enviroment, IDEA, Venezuela

Research focus

2 Author merit
Reviewer merit
Editor merit

Education

Bachelor, University Central of Venezuela, 2018

2 Protocols published

Isolation of PCR-quality Genomic DNA from Soils Impacted with Extra Heavy Crude Oil

Authors: Laynet A. Puentes , Yusibeska L. Ramos, Ysvic A. Inojosa, César E. Rivera and Angela De Sisto, date: 06/20/2019, view: 2472, Q&A: 0

For the study of microbial communities in samples of soils impacted with extra heavy crude oil, it is necessary to perform molecular analyses. Due to the difficulty of oil matrix handling, there are very few protocols reported in writing. Also, one ...

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Very Low Molecular Weight Proteins Electrophoresis Protocol

Authors: César E. Rivera, José D. Rosales, Juan C. Freites-Perez and Eliaira Rodriguez, date: 11/20/2018, view: 10191, Q&A: 4

The electrophoresis is the most used technique to separate proteins and usually the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) proposed by Laemmli is the prefered since it can diferentiate large size proteins, but for very ...

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vu hung

Immanuel Kant Baltic Federal University

Hello, thank you for your protocol. I'm interested in peptide 2 kDa. I have tried your protocol: 18% glycine - SDS- PAGE, 25 A for 3,5 hours. I use ruler 10- 250 kDa. The green band is 10kDa. And this is the results. Nearby the green band is 2 lanes of bromophenol (0,6 kDa as you wrote). I have 2 question:
- How can i reduce the time, because 3,5 hours is too long,
- How can i separate the geen band and bromophenol so that i can sure that i can detect peptide 2kDa between them.
Can i make 25% gel and put more ure??? Please reply me!!!! Thanks a lot

10/20/2020 6:48:52 AM Reply

vu hung

Immanuel Kant Baltic Federal University

Thank you for reply, can i ask 2 more questions:
- Why do we need to pre-run?
- in most of protocols, we need to run with samples at low Voltage so that protein can concentrate in stacking gel --- but don't we need this step for low molecular weight proteins???
Best regards,

2020-10-22 21:54:44

jose rosales

idea

Good morning, in the gel I see that the green band is very close to the end of the gel; which could indicate that the low molecular weight band came out of the gel during the electrophoretic run.

In the gel that I show the 10 kDa band is in the middle of the gel. Please check the acrylamide concentration of your gel.

You can use 20% acrylamide and 20 minutes of pre-run.

It does not change the urea concentration.

The amperage can be 30 mAmp, but take care of the temperature, that the buffer does not overheat, a cold room is useful.

2020-10-21 05:29:30