Biotin Conjugated Human HSPA1L Recombinant Rabbit Monoclonal Antibody [PSH29-29] - Detector

Sandwich ELISA analysis of Human HSPA1L protein matched pair antibodies

Capture: HA724857, Human HSPA1L Rabbit mAb [PSH29-28]
Detector: HA724858, Human HSPA1L Rabbit mAb [PSH29-29]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724857) diluted in carbonate/bicarbonate buffer, at a concentration of 2 μg/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 HSPA1L protein (HA211506) starting from 20,000 pg/mL to 0 pg/mL and detect antibody (HA724858, 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 HSPA1L in NCI-H1299 extract samples based on a 1,000 µg/mL extract load.

Capture: HA724857, Human HSPA1L Rabbit mAb [PSH29-28]
Detector: HA724858, Human HSPA1L Rabbit mAb [PSH29-29]

Interpolated concentration of native HSPA1L was measured in duplicate at different sample concentrations and interpolated from the HSPA1L standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean HSPA1L concentration was determined to be 3,053 pg/mL in NCI-H1299 cell extract.

Interpolated concentrations of spiked HSPA1L in cell culture media samples.

Capture: HA724857, Human HSPA1L Rabbit mAb [PSH29-28]
Detector: HA724858, Human HSPA1L Rabbit mAb [PSH29-29]

The concentrations of HSPA1L were measured in duplicates, interpolated from the HSPA1L 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).