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|Sample type||Average %||Range|
|Cell culture supernatant||98||96% - 100%|
|Serum||103||101% - 105%|
|Heparin Plasma||100||99% - 101%|
|EDTA Plasma||93||90% - 96%|
|Citrate Plasma||86||84% - 88%|
Abcam’s Human IL-1 beta SimpleStep ELISA® kit (ab184861) has been re-developed with new capture and detector antibodies. This new kit will have the same name but a different product number (ab214025). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantification of IL-1 beta in human serum, plasma, and cell culture supernatants.
IL-1beta in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-1beta protein in human serum, plasma and cell culture supernatants.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Interleukin 1 beta (IL-1 beta) is produced by activated macrophages and stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells.
|Components||1 x 96 tests|
|10X Human IL-1beta Capture Antibody||1 x 600µl|
|10X Human IL-1beta Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|ab221824 - Antibody Diluent 4BI||1 x 6ml|
|Human IL-1beta Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Sample Diluent NS||1 x 50ml|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab214025 in the following tested applications.
|Sandwich ELISA||Use at an assay dependent concentration.|
The concentrations of IL-1beta were measured in duplicates, interpolated from the IL-1beta standard curves and corrected for sample dilution. Undiluted samples are as follows: PBMC supernatant 50% and 100% THP-1 supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1beta concentration was determined to be 1301 pg/mL in PBMC supernatant and 734 pg/mL in THP-1 supernatant.
Interpolated values are plotted (mean +/- SD, n=2). IL-1beta was measured in 2 donor serum samples (30 pg/mL and 140 pg/mL) and the remaining 8 samples measured less than the lowest point of the IL-1beta standard curve.
Background-subtracted data values (mean +/- SD) are graphed.
Conditioned media was harvested after 48 hours. IL-1beta was measured in 50% unstimulated and PHA stimulated PBMC supernatant. The concentrations of IL-1beta were measured in duplicate, interpolated from the IL-1beta standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1beta concentration was determined to be 1273 pg/mL in PHA stimulated PBMC supernatant. There was no detectable signal in unstimulated supernatant.
Conditioned media was harvested after 48 hours. IL-1beta was measured in 100% unstimulated and LPS stimulated THP-1 supernatant. The concentrations of IL-1beta were measured in duplicate and interpolated from the IL-1beta standard curves. The interpolated values are plotted (mean +/- SD, n=2). The mean IL-1beta concentration was determined to be 718 pg/mL in LPS stimulated THP-1 supernatant. There was no detectable signal in unstimulated supernatant.
ab214025 has not yet been referenced specifically in any publications.