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|Sample type||Average %||Range|
|Urine||105||100% - 107%|
|Serum||109||106% - 110%|
|Cell culture media||104||102% - 107%|
|EDTA Plasma||107||87% - 120%|
|Citrate Plasma||109||98% - 116%|
|serum free media||99||98% - 103%|
Abcam’s Lipocalin-2 (NGAL) Mouse in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Lipocalin-2 protein in mouse serum, plasma, urine and cell culture supernatant.
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.
Lipocalin-2 (also known as Neutrophil gelatinase-associated lipocalin or NGAL) is an iron binding and iron trafficking protein. Lipocalin-2 is involved in multiple cellular processes including apoptosis, innate immunity and renal development. Mice deficient in Lipocalin-2 appear normal but have increased susceptibility to bacterial infection. The bacteriostatic function may be related to Lipocalin-2 limiting bacterial iron supply. Mouse Lipocalin-2 has 81% and 62% protein sequence identity to rat and human Lipocalin-2, respectively.
|Components||1 x 96 tests|
|10X Mouse Lipocalin-2 Capture Antibody||1 x 600µl|
|10X Mouse Lipocalin-2 Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|Antibody Diluent 5B||1 x 6ml|
|Mouse Lipocalin-2 Lyophilized Recombinant Protein||2 vials|
|Plate Seal||1 unit|
|Sample Diluent NS||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab199083 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Standard curve comparison between mouse Lipocalin-2 (NGAL) SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 7-fold increase in sensitivity.
Background-subtracted data values (mean +/- SD) are graphed.
The concentrations of Lipocalin-2 were measured in duplicate and interpolated from the Lipocalin-2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 265.2 ng/mL in mouse serum, 583.8 ng/mL in mouse plasma (Citrate), and 535.2 ng/mL in platelet poor mouse plasma (EDTA).
J774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. Raw data from duplicate measurements are plotted (mean +/- SD, n=2).
L929 and J774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. RAW264.7 were grown in the absence (unstimulated) or presence of LPS (stimulated) for 2 days. Cell Culture supernatant was diluted to within the range of the assay, 5-fold (L929 and J774A.1) or 2,000-fold (RAW264.7). Measured values were interpolated from the Lipocalin-2 Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed. Lipocalin-2 was undetectable in media.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"