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Mouse IL-6 ELISA MAX™
431304
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We used Mouse IL-6 ELISA MAX™ to measure iL-6 contents in serum of tumor-injected mice.
ELISA
Serum
(1) Overnight binding of Capture Antibody. (2) Wash plate 4 times. (3) Block non-specific binding with Assay Diluent for 1 hour at RT. (5) While plate is being blocked, prepare standard dilutions. (6) Wash plate 4 times with Wash Buffer. (7) Add standard dilutions and samples and incubate at RT for 2 hours. (8) Wash plate 4 times with Wash Buffer. (9) Add Detection Antibody and incubate at RT for 1 hour. (10) Wash plate 4 times with Wash Buffer. (11) Add Avidin-HRP solution for 30 minutes. (12) Wash plate 5 times with Wash Buffer. (13) Add TMB Substrate Solution and incubate in the dark for 15–30 minutes. (14) Stop reaction by adding of Stop Solution to each well. (15) Read absorbance at 450 nm within 15 minutes. If the reader can read at 570 nm, the absorbance at 570 nm can be subtracted from the absorbance at 450 nm.
Incubations should be done shaking on a plate shaker.
Mice with tumors had significantly higher amounts of IL-6 in serum.
N/A
The ELISA performed well
Low signal in the standards reduces sensitivity at low IL-6 concentrations.
This ELISA kit works well for routine measurement of IL-6 in mouse serum.