Indirect ELISA analysis of Anti-CXCL12 Recombinant Rabbit Polyclonal Antibody (Product # 711345) was performed by coating recombinant human CXCL12 protein standard at concentrations ranging from 0.24 ng/mL to 250 ng/mL onto a plate. The coated protein was bound by Anti-CXCL12 Recombinant Rabbit Polyclonal Antibody at 1 µg/mL and was detected using Goat anti-Rabbit IgG (H+L) secondary antibody, HRP-conjugate (Product # G-21234, 1:5000 dilution). The plate was developed using stabilized chromogen solution, TMB+H2O2 (Product # SB01). The plate was read at 450 nm with Thermo Scientific Multiskan GO microplate spectrophotometer. The endogenous levels of CXCL12 in diseased human serum (Atopic dermatitis) and plasma (pooled atopic dermatitis, obesity and rheumatoid arthritis) along with simulated cell treatment (THP1+LPS @ 1 µg/mL for 24 hrs+PMA @ 100 nM for 10 mins) was quantitated by an indirect ELISA analysis by coating the samples onto the plate as per dilutions mentioned. The samples were detected as mentioned above. Note : The experiment was performed using an antibody pair buffer kit (Product # CNB0011).