see above
I am currently culturing the T-CLL, Kit-225 cell line. My culture condition is RPMI 10% FCS with 20ng/ml IL2 (R&D, about 420IU/ml). Cells look viable but proliferation rate is slow. Is there...
05 June 2017 6,536 6 View
I am facing problem in qPCR analysis. The file attached shows the melt curve of GAPDH. In my setting, my template was 100ng and primer concentration 500nM in 20ul. Cycling number was 40 cycles. In...
31 December 2015 5,756 5 View
I am currently comparing ULBP-4 expression in cancer cells. The method I used for qPCR as follow: a) cDNA synthesis = 1ug of RNA in 20ul b) I diluted the cDNA 10X prior to qPCR c) I used 4ul of...
09 October 2015 6,358 9 View
I have run 3 times of RT-PCR for one of my sample. However, fold changes always inconsistant in these three attempts. What is the possible factors contribute to this inconsistancy?
06 July 2015 6,622 5 View
I am currently doing western blot for p-Akt. Below is my protocol: -- Blocking = RT, 2 hours with 5% BSA -- Primary antibody = p-Akt (1:2000), overnight, RT -- Secondary antibody = Anti rabbit...
04 May 2015 6,233 9 View
Some protocols mention primary antibodies should be diluted in 5%BSA whereas some protocols mention primary antibodies should be diluted in 5% non-fat milk. What is the difference? What is the...
04 May 2014 479 20 View
Many publications mention that cells were serum starved for 24 hours before adding TGF-beta or EGF to induce EMT. After serum starved, is it possible to treat cells with EMT-inducers in RPMI 5%...
04 May 2014 7,810 10 View
I have found that cbioportal is a good website to study molecular expression level of certain proteins in certain cancer cell lines. Unfortunately, some cell lines are not in the database. Where...
03 April 2014 1,073 4 View
The supplier COA stated that the snail protein is about 30KD. I ran the electrophoresis at 200V, 125mA for 45mins. I used 4-12% Tris Nu-PAge Gel. Why is the snail protein more than 30KD?
02 March 2014 8,151 7 View
I am currently comparing snail-1 expression in esophageal cancer cell lines. My protein load was 30-40 ug, the primary snail-1 antibody (cell signaling) was 1:1000, and the secondary anti-rabbit...
02 March 2014 8,731 17 View
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
I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...
07 August 2024 6,034 3 View
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
Evolutionary fitness is based on an organism’s ability to adapt rapidly to changing environmental circumstances. Large-bodied mammals have been equipped with large brains (and hence a high...
06 August 2024 4,849 2 View
Air moisture harvesting Air water collection devices
06 August 2024 5,473 2 View
i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 View
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
Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...
04 August 2024 8,894 1 View
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
what are the top 3 challenges to the advancement of the field of Radiogenomics in cancer research? is it the availability of easily available low-cost matched imaging and biosamples with clinical...
03 August 2024 5,828 4 View
