Biotin Conjugated Human/Monkey TIMP4 Recombinant Rabbit Monoclonal Antibody [PSH21-50] - Detector

Sandwich ELISA analysis of Human TIMP4 matched pair antibodies

Capture: HA724220, Human/Monkey TIMP4 Rabbit mAb [PSH21-49]
Detector: HA724221, Human/Monkey TIMP4 Rabbit mAb [PSH21-50]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724220) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human TIMP4 protein (HA211288) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA724221, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Interpolated concentrations of native TIMP-4 in human and monkey samples.

Capture: HA724220, Human/Monkey TIMP4 Rabbit mAb [PSH21-49]
Detector: HA724221, Human/Monkey TIMP4 Rabbit mAb [PSH21-50]

Interpolated concentration of native TIMP-4 was measured in duplicate at different sample concentrations and interpolated from the TIMP-4 standard curves. Undiluted samples were 100% serum. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean TIMP-4 concentration was determined to be 2,043 pg/mL in human serum and 1,031 pg/mL in monkey serum.

Interpolated concentrations of spiked TIMP-4 in cell culture media samples.

Capture: HA724220, Human/Monkey TIMP4 Rabbit mAb [PSH21-49]
Detector: HA724221, Human/Monkey TIMP4 Rabbit mAb [PSH21-50]

The concentrations of TIMP-4 were measured in duplicates, interpolated from the TIMP-4 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).