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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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
hematopoietin-1; IL-1 alpha; interleukin 1, alpha; interleukin-1 alpha; preinterleukin 1 alpha; pro-interleukin-1-alpha
IL-1A; IL1; IL1-ALPHA; IL1A; IL1F1