Quynh Pham

8 Questions 8 Answers 0 Followers

Questions related from Quynh Pham

Why does my protein bind poorly to the Ni sepharose beads?

I am purifying RNAse E in E. coli (118 kDa). I cloned the rne coding sequence on pET28a with a His tag at the N-terminus. The protein overexpressed fine (see attached figures). The binding buffer...

25 June 2024 6,162 4 View

Why couldn't my protein bind onto the membrane in dot blot?

I tried to detect my recombinant protein using dot blot. After spotting the cell lysate onto the membrane, I let it dry at room temperature for 1 hour, and then proceeded with blocking and...

11 January 2024 4,233 5 View

How can you achieve successful Western blot results for phosphorylated proteins?

I am trying to detect the phosphorylated IKK protein using Western Blot. I induced the cells by 10ug/ml LPS for 20 min. I loaded approximately 25 μg of total lysate and ran the gel. After gel...

17 November 2023 6,299 3 View

Why is my Urea PAGE 10% gel unable to separate the RNAs well?

I performed the RNase footprint with an 106-nt RNA. However, my hydrolysis ladder showed poor resolution from the >35-nt bands (attached figure). I used the 8M Urea gel with 10% AcrylamideBis...

09 February 2022 6,138 0 View

Why did my protein gel cannot be stained with silver staining properly?

I am doing silver staining since my protein concentration is low. However, the gel became dark and the protein bands appear as negative band (even the bands which are not present in coomassie...

21 June 2018 2,066 2 View

Using Nanodrop to estimate protein concentration shows high peak at 260 nm absorption?

I am using Nanodrop to measure protein concentration in lysate since it is fast and only requires small amount of sample. When I use water as blank, the result showed two high peak at 230 and 280...

08 November 2017 8,164 6 View

How many samples should we process to extract RNA at the same time?

I am using kits from Thermorscientific to extract total RNA. I extracted more than 20 samples and the results were various. Some of them had high concentration and the other ones were low. I do...

16 March 2015 9,489 4 View

Can RNA be degraded in -20°C after 24 hours?

I extracted RNA by GeneJET Plant RNA Purification Mini Kit from Thermoscientific and got 4 bands in undenatured electrophoresis (without DNAse treatment). Then I stored it in -20 °C. After about...

29 October 2014 3,775 9 View