Sandwich ELISA analysis of human IL-1 alpha was performed using a Human IL-1 alpha Colorimetric ELISA Kit (Product # EH2IL1A) by loading 50ul per well of Human IL-1 alpha Recombinant Protein (Product # SIL1A) in dodecuplicate at 400, 160, 64, 25.6, 10.25 and 0 pg/ml across a 3ug/ml mouse anti-human IL-1 alpha (Product # M421A) pre-coated plate and incubating for 1 hour at room temperature. The plate was washed, then incubated with 50ul per well of a biotinylated-mouse anti-human IL-1 alpha (Product # M420AB) in dodecuplicate at 0.15µg/ml for 1 hour at room temperature. The plate was washed and incubated with 100ul per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at 1:10 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant human IL-1 alpha|
|Storage buffer||PBS with 4% BSA|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
M420AB targets IL-1 alpha in ELISA applications and shows reactivity with Human samples.
The M420AB immunogen is recombinant human IL-1 alpha.
M420AB detects IL-1 alpha which has a predicted molecular weight of approximately 18 kDa.
The M420AB IL1a antibody (biotinylated conjugate of clone B133.5) has successfully been paired as the detection antibody in a sandwich ELISA with capture antibody M700A (clone 2G1). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Biotinylated antibody M420AB (clone ILA8-H12) and antibody M421A (clone ILA9-H18) have successfully been used in combination with recombinant IL1 alpha protein SIL1A in ELISA applications.
The protein encoded by this gene is a member of the interleukin 1 cytokine family. This cytokine is a pleiotropic cytokine involved in various immune responses, inflammatory processes, and hematopoiesis. This cytokine 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.
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