Yang Zhang

7 Questions 4 Answers 0 Followers

Questions related from Yang Zhang

How to do DNA normalization based on qPCR values?

Hi Everyone: I have a question regarding to qPCR. I am trying to do RT-PCR, and I want to have equal amount of input DNA for my RT-PCR, so I normalize my DNA input based on qPCR values of a...

01 February 2021 7,199 4 View

MS/MS protein analysis?

Hi Everyone: I want to quantify the relative amount of a protein in 2 samples. I will first digest the sample with trypsin, and subject the resulting peptides to LC-MS/MS analysis. I will then...

26 January 2021 8,547 3 View

How do I normalize RT-qPCR results?

Hi Everyone: I am studying a protein that is hypothesized to assist in transcription elongation. Its knockdown is expected to lead to decreased RNAPII processivity. I am trying to test this...

12 February 2020 556 4 View

DNA ethanoal precipitation contamination?

Hi all, I use Q5 NEB high-fidelity polymerase for PCR. I usually do 4 replicates, and ethanol precipitate the PCR products (0.1 sample volume 3 M NaAc, 2 sample volume of ethanol). I got a big...

28 September 2019 2,742 6 View

Protein treated with Calf intestinal phosphatase has increased mobility on gel. Does that indicate the protein is phosphorylated?

I have a protein of interest. Let's call it protein X. I treat the cell extract with calf intestinal phosphatase (from NEB). incubate at 37 degree for 1 h. Protein X would have increased mobility...

22 April 2019 7,952 3 View

Pol II CTD phosphorylation detection

Hi All, Pol II RPB1 CTD is phosphorylated, and can be detected as doublet bands on gel. I expressed a Myc tagged Rpb1 in cell, and tried to detect Rpb1-Myc. However, I can only see one band with...

01 January 1970 1,813 4 View

How to probe protein phosphorylation without an antibody?

Hi All, I have a protein of interest, X, and it has a single phosphorylation site identified. Another protein A might dephosphorylate X. I want to knock down A, and quantify the change of...

01 January 1970 9,691 7 View