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|Cell culture supernatant||88.8||87.4% - 90.3%|
|Serum||101.2||98.5% - 103.5%|
|Cell culture media||94.4||93.4% - 96.1%|
|Citrate Plasma||97.6||94.1% - 101.6%|
IL-1a (Interleukin-1 alpha) mouse in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-1a in mouse serum, plasma (citrate), and cell culture supernatant samples.
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 (IL-1) refers to two proteins encoded by two different genes (IL-1a and IL-1b), both of which share the same cell surface receptors. Mouse IL-1a is a pro-inflammatory cytokine that encodes a 270 aa residue called pro-IL-1a, which contains a nuclear localization sequence, an N-myristoylation site, and three potential N-linked glycosylation sites. Primarily, pro-IL-1a is localized to the cytosol after synthesis. However, some pro-IL-1a is secreted and is subsequently cleaved by extracellular proteases such as calpain to form the 17 kDa, 154 aa residue mature IL-1a. Importantly, both pro-IL-1a and mature IL-1a have been shown to bind the IL-1 receptor and are biologically active. This is different than pro-IL-1b and mature IL-1b, where pro-IL-1b is biologically inactive. Within the mature protein, rat IL-1a and human IL-1a share 79% and 56% aa sequence identity, respectively, with the mouse IL-1a protein.
IL-1a and IL-1b interact with three type I transmembrane Ig superfamily proteins: IL-1 receptor type I (IL-1 RI), IL-1 RII, and IL-1 receptor accessory protein (IL-1 RAcP). Both IL-1 RI and IL-1 RII can bind directly to IL-1a and IL-1b. IL-1 RAcP interacts with IL-1 RI in the presence of IL-1 to form a high-affinity receptor complex that is required for intracellular signal transduction. Interestingly, IL-1 RAcP will also bind IL-1 RII, but this creates a non-functional high-affinity receptor complex that does not transduce IL-1 signals. Thus, IL-1 RII can act to moderate the function of IL-1a and IL-1b.
IL-1a is typically expressed under normal conditions in skin keratinocytes and some epithelial cells, as well as in some cells in the central nervous system. Under pathological conditions, IL-1a is also strongly expressed by monocytes, tissue macrophages, dendritic cells, B lymphocytes and NK cells. Intracellular pro-IL-1a can also be localized to the nucleus where it plays a role as an intracellular regulator of human endothelial cell proliferation and migration. IL-1 possesses a wide variety of biological activities and plays a central role in mediating immune and inflammatory responses. Normal production of IL-1 is critical for hematopoiesis, osteoclast differentiation, and initiation of normal host responses to stresses such as injury and infection. Incorrect production of IL-1 has been shown to be associated with a variety of pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin-dependent diabetes mellitus, and atherosclerosis.
|内容||1 x 96 tests|
|10X Mouse IL-1a (Interleukin-1 alpha) Capture Antibody||1 x 600µl|
|10X Mouse IL-1a (Interleukin-1 alpha) Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|Antibody Diluent CPI - HAMA Blocker (ab193969)||1 x 6ml|
|Mouse IL-1a (Interleukin-1 alpha) Lyophilized Recombinant Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS (ab193972)||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 ab199076 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 IL-1a SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.
Background-subtracted data values (mean +/- SD) are graphed.
Recombinant IL-1a mouse was spiked into 50% mouse serum and 50% mouse plasma and diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse IL-1a were measured in duplicate and interpolated from the mouse IL-1a standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).
RAW264.7 cells were cultured in high glucose DMEM with 10% fetal calf serum or horse serum, 2 mM L-glutamine and 100 µg/mL Kanamycin. RAW264.7 cells were starved for 24 hours and treated in the presence and absence of 5 µg/mL of LPS. The concentrations of IL1-a were interpolated from the calibration curve and corrected for sample dilution (1:2). The mean IL-1a concentration in RAW264.7 media was determined to be 288 pg/mL in stimulated RAW 264.7 cells and undetectable in unstimulated RAW264.7 cells.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"