I'm going to do Western Blot and I would like to know what is the best kit to use to extract the protein from adipocytes cell (3T3-L1)?
You may use M-PER (Mammalian-Protein Extraction Reagent) from thermo-pierce. Best...
MPER with Protease inhibitors works fine. I did in hepatic cells. Extraction was fine!!
I'm running Real-time PCR on STEPONEplus instrument (singleplex) which allows me to choose only one endogenous gene. However, can I run another endogenous gene in the same 96 well? I would also...
04 May 2013 2,448 2 View
Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...
11 August 2024 7,082 3 View
I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...
10 August 2024 7,480 3 View
I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...
08 August 2024 9,882 2 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...
05 August 2024 9,798 3 View
I will be with my students collecting seaweed samples in a marine farm and later we will process this tissue for RNA isolation and further sequencing. Does anyone have tips on how to collect the...
04 August 2024 501 2 View
HOW CAN I WRITE A CODE TO USE THE WAVENET TRANSFORM AS A FEATURE EXTRACTION METHOD INSTEAD OF DWT IN MATLAB?
03 August 2024 7,829 0 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
I am currently working on a project that involves extracting cell membranes, for which we disrupt the cells using sonication. During the initial extraction process, we add protease inhibitors to...
30 July 2024 7,077 1 View
Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....
29 July 2024 5,007 0 View