製品の概要

  • 製品名
  • 検出方法
    Colorimetric
  • 再現性
    Intra-Assay(同時再現性)
    サンプル N 平均値 SD CV%
    Overall 8 2.31%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    Overall 3 4.78%
  • サンプルの種類
    Cell culture supernatant, Serum, Citrate Plasma
  • アッセイタイプ
    Sandwich (quantitative)
  • 検出感度
    0.62 pg/ml
  • 検出範囲
    4.69 pg/ml - 300 pg/ml
  • 添加回収試験

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    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%

  • 全工程の試験時間
    1h 30m
  • ステップ
    One step assay
  • 種交差性
    交差種: Mouse
  • 製品の概要

    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

  • 特記事項

    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.

  • アプリケーション
    適用あり: Sandwich ELISAmore details
  • 試験プラットフォーム
    Microplate (12 x 8 well strips)

製品の特性

  • 保存方法
    Store at +4°C. Please refer to protocols.
  • 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 Development Solution 1 x 12ml
  • 研究分野
  • 関連性
    IL-1 alpha is a member of the interleukin 1 cytokine family. It is a pleiotropic cytokine involved in various immune responses, inflammatory processes, and hematopoiesis. IL-1 alpha is produced by monocytes and macrophages as a proprotein, which is proteolytically processed and released in response to cell injury, and thus induces apoptosis. This gene and eight other interleukin 1 family genes form a cytokine gene cluster on chromosome 2. It has been suggested that the polymorphism of these genes is associated with rheumatoid arthritis and Alzheimer's disease.
  • 細胞内局在
    Secreted
  • 別名
    • Hematopoietin 1
    • IL 1 alpha
    • Il 1a
    • IL1
    • IL1 ALPHA
    • IL1A
    • IL1F1
    • Ilia
    • Interleukin 1 alpha
    • Interleukin 1 alpha precursor
    • Interleukin1 alpha
    • OTTHUMP00000203692
    • OTTMUSP00000016430
    • Preinterleukin 1
    • Pro interleukin 1
    • Pro interleukin 1 alpha
    • Prointerleukin 1
    • RP23-160G19.8
    see all
  • 参照データベース

関連製品

アプリケーション

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.

アプリケーション Abreviews 特記事項
Sandwich ELISA Use at an assay dependent concentration.

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  • 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.

プロトコール

参考文献

This product has been referenced in:
  • Ren LR  et al. Staphylococcus Aureus Induces Osteoclastogenesis via the NF-?B Signaling Pathway. Med Sci Monit 23:4579-4590 (2017). Read more (PubMed: 28942456) »
See 1 Publication for this product

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