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%
    PBMC PHA+ 24 3.9%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    PBMC PHA+ 3 4.7%
  • サンプルの種類

    Cell culture supernatant, Serum, Plasma
  • アッセイタイプ

    Sandwich (quantitative)
  • 検出感度

    0.5 pg/ml
  • 検出範囲

    3.1 pg/ml - 200 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
    Serum 104 103% - 108%
    Cell culture media 95 91% - 99%
    Hep Plasma 104 100% - 108%
    EDTA Plasma 102 91% - 120%
    Cit plasma 103 88% - 119%

  • 全工程の試験時間

    1h 30m
  • ステップ

    One step assay
  • 種交差性

    交差種: Human
  • 製品の概要

    Human IL-1a SimpleStep ELISA® kit has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency. This version will be discontinued March 2020 unless inventory is depleted beforehand.


    IL-1a (Interleukin-1 alpha) in vitro SimpleStep ELISA® kit is designed for the quantitative measurement of IL-1a protein in human serum, plasma and culture supernatants.


    Note: Normal human serum IL-1a levels are below the detection limit of this kit.

    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. IL-1a is a proinflammatory cytokine typically expressed under normal conditions in skin keratinocytes, some epithelial cells as well as 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. IL-1a is synthesized as a 31kDa precursor, some of which is cleaved prior to secretion into a 17.5kDa protein. IL-1a is biologically active both in a paracrine or systemic mode of action. The paracrine effects are mediated by the uncleaved portion of IL-1a, which is transported to the cell surface where it activates the IL-1 receptor type I (IL-1RI) of adjacent cells. The systemic effect is mediated by cleaved IL-1a, which is secreted into the bloodstream.

    Signaling of IL-1 via its receptor, results in recruitment of the IL-1 R accessory protein (IL-1RAcP). This leads to a series of phosphorylation and ubiquitination events that ultimately activate the nuclear factor kB (NF-kB) and signal via the c-Jun N-terminal kinase (JNK) and the p38 mitogen-activated protein kinase (MAPK) pathways. These signaling events generate local and systemic responses that mediate acute phase inflammation, a critical host response to injury and infection. Under these circumstances, IL-1 induces fever, pain sensitivity, vasodilation, hypotention and slow wave sleep. All these responses are essential processes geared towards the re-establishment of tissue homeostasis. However, prolonged and inappropriate IL-1 induction has been shown to be associated with 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

製品の特性

  • 保存方法

    Store at +4°C. Please refer to protocols.
  • 内容 1 x 96 tests
    10X Human IL-1a Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Human IL-1a Capture Antibody (Lyophilized) 2 vials
    Human IL-1a Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent 25BS 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
  • 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
    • IL 1 alpha
    • Il 1a
    • IL1
    • IL1 ALPHA
    • IL1-a
    • IL1A
    • il1alpha
    • IL1F1
    • Ilia
    • Interleukin 1 alpha
    • Interleukin 1 alpha precursor
    • interleukin 1a
    • 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

アプリケーション

Our Abpromise guarantee covers the use of ab178008 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.

画像

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Human PBMC 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 37ºC in the presence or absence of PHA. The concentrations of IL-1a were interpolated from the calibration curve and corrected for sample dilution. The mean IL-1a concentration was determined to be 1.4 pg/mL in unstimulated PBMC supernatants and 96 pg/mL in stimulated PBMCs.

プロトコール

参考文献

ab178008 has not yet been referenced specifically in any publications.

レビューと Q&A

Answer

Thank you for contacting us.



Only the cleaved/mature form is secreted, therefore we are confident that the assay measures cleaved IL-1a as we can measure levels of IL1a in stimulated PBMCs in comparison to the non-stimulated PBMCs supernatant. We have not tested whether the assay measures non-cleaved IL1a.

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