Robinson White

10 Questions 8 Answers 0 Followers

Questions related from Robinson White

How to interpret the following abnormal dual luciferase assay result?

I have transfected Hep G2 cell with my promoter of interest having pGL3 backbone and the pRL-TK vector as internal control using lipofectamine ltx following the company protocol. After 24 h...

13 July 2017 3,973 0 View

Long time effect of cell culture medium on adherent or suspension cell

I am working with HL-60 (suspension cell) and HuH7 (adherent cell). Normally it is seen that the cells become confluent within 3 days after subculture; while sometimes it does not. It may take 1...

02 July 2017 5,612 3 View

THP-1 differentiation and transfection- easy or difficult?

I would like to differentiate THP-1 cell to macrophage using PMA and transfect pAcGFP vector with Lipofectamine ltx transfection reagent (as it is available). Is there any standard protocol of...

15 June 2017 4,279 2 View

How to find binding sites of transcription factors from the nucleotide sequence of promoter of interest-reporter vector clone?

I have got a sequence of my human promoter of interest-reporter (pGL4) vector of 3Kbp. The promoter sequence is in capital letters (representing the nucleotides) and the reporter vector sequence...

29 March 2017 4,668 4 View

How to interpret dual luciferase assay result (RLU) and calculate transfection efficiency?

I performed a dual luciferase assay using pGL3 firefly luc as experimental reporter and pRL-TK renilla luc as internal control (Promega assay kit). I used a manual luminometer to analyze my...

25 March 2017 9,528 8 View

Standard protocol for the Dual Luciferase Assay with HL-60 cell line

I want to perform the dual luc. assay using Hl-60 cell line which is a floating cell. Therefore, I explored different papers but no brief protocol found about the assay using this floating...

15 March 2017 9,024 1 View

What are the reasons for getting extra bands after electrophoresis?

I run 0.7% agarose gel electrophoresis of my plasmid DNA sample consisting of GFP vector using NheI and HindIII restriction enzyme. I used two types of plasmids, i.e., isolated by mini prep-alkali...

06 January 2017 7,437 1 View

How to prevent HL-60 cell contamination?

I have thawed Hl-60 cells from liquid nitrogen chamber two times in RPMI 1640 media (containing 10% FBS, L glutamin, penicillin/streptomycin). In the first day the cells remain okay. But when...

07 December 2016 3,541 7 View

How easily transfect the floating cell, HL-60 using X-tremeGENE HP DNA Transfection reagent?

I need to transfect GFP vector and Luciferase vector plasmid DNA into the HL-60 floating cells. I will have to use X-tremeGENE HP DNA Transfection reagent from ROCHE. If anyone already experienced...

24 November 2016 5,616 2 View

Why phosphorylated (p)-ERK 1/2 down regulated soon after activation?

Generally, we know that ERK signaling is responsible for controlling cell growth, proliferation, differentiation, and apoptosis. However, I found this signal important for cytokine expression in...

01 January 1970 6,649 3 View