TNF-α Autocrine Feedback Loops in Human Monocytes: The Pro- and Anti-Inflammatory Roles of the TNF-α Receptors Support the Concept of Selective TNFR1 Blockade In Vivo

Table 1

Assays.

mAb	Clone number	Fluorophore	Function	Manufacturer
Mouse IgG1 TNF-α	28401	N/A	Blocking	R and D Systems
Mouse IgG1 TNFRI	16805	N/A	Blocking	R and D Systems
Mouse IgG1 TNFR2	22210	N/A	Blocking	R and D Systems
Mouse IgG1 isotype control	11711	N/A	Control	R and D Systems
Mouse IgG1 TNFR1	16803	PE	Flow cytometry	R and D Systems
Mouse IgG TNFR2	22235	FITC	Flow cytometry	R and D Systems
Mouse IgG1 isotype control	11711	PE	Flow cytometry	R and D Systems
Mouse IgG control	X39	FITC	Flow cytometry	BD Biosciences
ELISA	Minimum detectable dose (pg/mL)	Lower limit of quantification (pg/mL)	Upper limit of quantification (pg/mL)	Manufacturer
TNF-α	1.6	15.6	1000	R and D Systems
CXCL8	3.5	31.3	1000	R and D Systems
TNFR1	50	50	45000	Invitrogen
TNFR2	100	100	138000	Invitrogen
Gene symbol (TaqMan™)	Assay-on-demand number	NCBI RefSeq transcript number	Amplicon length	Manufacturer
GAPDH	Hs99999905_m1	NM_002046.5	122 bp	Life Technologies
TNF-α	Hs00174128_m1	NM_000594.3	80 bp	Life Technologies
CXCL8	Hs00174103_m1	NM_000584.3	101 bp	Life Technologies
IL-1β	Hs01555410_m1	NM_000576.2	91 bp	Life Technologies
IL-6	Hs00985639_m1	NM_000600.3	66 bp	Life Technologies
TGF-β	Hs00998133_m1	NM_000660.4	57 bp	Life Technologies
IL-10	Hs00961622_m1	NM_000572.2	74 bp	Life Technologies

Details of blocking and flow cytometry antibodies. ELISAs and TaqMan gene expression assays are supplied.