TNF-α determinations, IL-6, IL-10 and nitrite / nitrate (NOx) routine employed as a surrogate marker of NO metabolism in mouse plasma by ELISA (commercially available kits) ratio were performed.
Both RNAzol RT and RNeasy Plant Mini kit produced A260/280 ratio at the range of 1.70 1.80 while a modified CTAB method produced the ratio at the range of 2.00 2.20.
Whole heart cell lysates (n = 3) were analyzed using a Bioassay Systems Enzylight™ ATP ADP ratio kit (ELDT-100, Bioassay Systems, Hayward, California).
After adding phosphate buffered saline with Tween buffer (PBST, provided in the kit) at a ratio of 1∶10 (sample material f.w.∶buffer) and a 5 mm tungsten carbide bead, leaf samples were macerated for 100 sec at 15 Hz and aphid samples for 40 sec at 30 Hz, using a mixer mill MM300 (Retsch, Haan, Germany) fitted with 24 tube-adapters (Qiagen, Hombrechtikon, Switzerland).
Using cell lines established from the spleen of various diseased mice, we observed that each of these cell lines exhibited a particular growth rate in response to either Epo alone or SCF alone, and expressed EpoR and Kit in a ratio modulated by the cytokine used to sustain their proliferation.
ADP/ATP ratios were determined using the fluorescent ApoSensor ADP/ATP Ratio kit (BioVision).
ADP and ATP levels were measured using an ADP/ATP ratio kit (Abcam).
Only embryos extracted via the Plus Universal, Maxwell® and SimplyRT HT kits and blood extracted via the Simply RT HT kit had A260 A280 ratios <1.8 and these less-than-optimal ratios were likely related to low concentrations (<15 ng/μL in all cases) of nucleic acid in these samples [ 24], rather than the ability of the kit to remove contaminants that absorb at 280 nm.
This in combination with the fact that half of the blood samples extracted via this kit had A260 A280 ratios <1.8 indicates that the SimplyRNA HT kit lacked precision in isolating "pure" RNA from blood samples.
ADP/ATP ratios were determined using the EnzyLight™ ADP/ATP ratio assay kit from BioAssay Systems (Hayward, CA, USA) according to the manufacturer's instructions.
