I'm seeing cell death after transfection using Lipofectamine RNAiMAX Reagent. What should I do?
Remove antibiotics from the medium during transfection, as this can cause stress leading to cell death. Try adjusting both lipid and siRNA quantities. Use cells with lower passage number for better viability. Consider assaying for microbes such as Mycoplasma that are not detected easily in cell culture.
I accidentally froze my Lipofectamine RNAiMAX Reagent. Can I still use it?
Freezing can alter the integrity and composition of the lipid particles. The performance of the reagent may be affected and it may only work for the first week or so. It is safer to use a new vial.
What positive transfection control can I use with Lipofectamine RNAiMAX?
BLOCK-iT Alexa Fluor Red Fluorescent Control (Cat. No. 14750100) can be used as an indicator of transfection efficiency of siRNA. This dye can be detected using standard filter sets designed for Alexa Fluor 555, Cy3, DsRed, Texas Red, or Rhodamine Red fluorophores.
BRAG2/GEP100/IQSec1 Interacts with Clathrin and Regulates a5ß1 Integrin Endocytosis through Activation of ADP Ribosylation Factor 5 (Arf5).
Authors:Moravec R, Conger KK, D'Souza R, Allison AB, Casanova JE,
Journal:J Biol Chem
PubMed ID:22815487
ADP ribosylation factors (Arfs) are small GTP-binding proteins known for their role in vesicular transport, where they nucleate the assembly of coat protein complexes at sites of carrier vesicle formation. Similar to other GTPases, Arfs require guanine nucleotide exchange factors to catalyze GTP loading and activation. One subfamily of ArfGEFs, ... More
Low-density Lipoprotein Receptor-related Protein-1 (LRP1) Mediates Autophagy and Apoptosis Caused by Helicobacter pylori VacA.
Authors:Yahiro K, Satoh M, Nakano M, Hisatsune J, Isomoto H, Sap J, Suzuki H, Nomura F, Noda M, Moss J, Hirayama T,
Journal:J Biol Chem
PubMed ID:22822085
In Helicobacter pylori infection, vacuolating cytotoxin (VacA)-induced mitochondrial damage leading to apoptosis is believed to be a major cause of cell death. It has also been proposed that VacA-induced autophagy serves as a host mechanism to limit toxin-induced cellular damage. Apoptosis and autophagy are two dynamic and opposing processes that ... More