Peripheral blood leukocytes of three human donors were isolated from blood bank buffy-coat preparations by Ficoll-Hypaque density gradient centrifugation, and T-cells were purified with high affinity negative selection human CD3+ T-cell enrichment columns. The cells were resuspended in RPMI-1640 with 10% FCS with approximately 3-8 millions cells/mL at room temperature overnight. T-cells were then either unactivated (1g 0 hr), activated (1g 4 hr) with a final concentration of 5 mg/mL Con A + 4 mg/mL anti-CD28 antibody and incubated for 4 hours at 37°C, or loaded onto a RPM rotating at 60°/s. T-cells loaded on the RPM were cultured for 2 hours prior to collection or activation to allow equilibration of cells to the new environment. After incubation, 1 mL of 4 M guanidinium isothiocyanate in 750 mM sodium citrate buffer, pH 7, with N-lauroylsarcosine and ß-mercaptoethanol was added to lyse cells and stabilize RNA. Lysates were frozen at -80°C. Lot batch = 2001996
The number of probe pairs in the probe set used to assign a detection call.
VALUE
A median normalized quantitative measure of the relative abundance of a transcript.
ABS_CALL
A qualitative measurement indicating if the transcript is detected (Present), not detected (Absent), or marginally detected (Marginal).
DETECTION P-VALUE
A p-value indicating the significance of the Detection call. A Detection p-value measures the probability that the discrimination scores of all probe pairs in the probe set are above a threshold and that the target transcript is likely to be present.
