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|Sample type||Average %||Range|
|Serum||101.3||89.9% - 110.09%|
|Cell culture media||106.57||97.09% - 116.04%|
|Heparin Plasma||100.07||87.61% - 116.98%|
|EDTA Plasma||96.92||94.06% - 100.01%|
Abcam’s MIP2 (CXCL2) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MIP2 (CXCL2) 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.
Macrophage inflammatory protein 2 (MIP2) otherwise known as CXCL2 GRO-beta| or Hematopoietic synergistic factor is a 7.9 kDa heparin-binding chemokine that has potent effects in the response to inflammation and induction of peripheral tolerance. MIP2 shares 70% sequence similarity with mouse and rat MIP2. It is secreted by activated monocytes neutrophils and inflamed mucosal epithelial cells in response to inflammatory stimuli such as IL-1β. MIP2 recruits granulocytic neutrophils and macrophages at sights of inflammation| and causes degranulation of these effector cells at the inflammatory site. It has also been hypothesized that MIP2 acts to synergize the effects of Granulocyte macrophage colony-stimulating factor (GM-CSF) and Macrophage colony-stimulating factor (M-CSF) leading to a larger recruitment of neutrophils and macrophages at the site of inflammation. Research has also suggested that MIP2 is involved in tolerogenic pathways as a lack of the MIP2 receptors CXCR1 and CXCR2 are associated with peripheral tolerance. Clinically. MIP2 is being used as a bone marrow transplant regimen to mobilize peripheral blood hematopoietic stem cells (PBSCs). Mobilization by MIP2 therapy in combination with G-CSF has been associated with faster immune cell reconstitution after transplantation.
|Components||1 x 96 tests|
|10X MIP2 Capture Antibody||1 x 600µl|
|10X MIP2 Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|ab221824 - Antibody Diluent 4BI||1 x 6ml|
|MIP2 Human Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent 25BP||1 x 20ml|
|Sample Diluent NS (ab193972)||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab184862 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.|
Example of MIP2 standard curve in Sample Diluent NS using ab184862
Interpolated values corrected by dilution factor (mean +/- SD) are graphed.
Example of MIP2 standard curve in Sample Diluent 25BP using ab184862
Human PBMCs were cultured in RPMI supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin. Cells were cultured for 2 days at 37˚C in the presence or absence of PHA. The concentrations of MIP2 were interpolated from the calibration curve and corrected for sample dilution. The mean MIP2 concentration was 67 pg/mL on unstimulated PBMC supernatants and 773 pg/mL on stimulated PBMCs supernatants.
Human MIP2 levels in individual healthy donors. Ten individual healthy donors were evaluated for the presence of MIP2 in serum using this assay. Results were interpolated from the standard curve in Sample Diluent 25BP and corrected for sample dilution (1:4). The mean level of Human MIP2 was found at 84.757 pg/mL with a range of 26.767 – 266.256 pg/mL.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"