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|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant human IL-1 alpha|
|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.
M421A targets IL-1 alpha in ELISA applications and shows reactivity with Human samples.
The M421A immunogen is recombinant human IL-1 alpha.
M421A detects IL-1 alpha which has a predicted molecular weight of approximately 18 kDa.
The M421A IL1a antibody (clone ILA9-H18) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M420AB (biotinylated conjugate of clone ILA8-H12). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody M421A (clone ILA9-H18) and biotinylated antibody M420AB (clone ILA8-H12) 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.
Bioactive interleukin-1alpha is cytolytically released from Candida albicans-infected oral epithelial cells.
M421A was used in immunoprecipitation to study IL-1 alpha release by Candida albicans-infected oral epithelial cells
|Dongari-Bagtzoglou A,Kashleva H,Villar CC||Medical mycology (42:531)||2004|
Prostaglandin E2 induces the final maturation of IL-12-deficient CD1a+CD83+ dendritic cells: the levels of IL-12 are determined during the final dendritic cell maturation and are resistant to further modulation.
M421A was used in ELISA to investigate the roles of prostaglandin E2 and interleukin 10 in CD1a+ dendritic cell maturation
|Kali¿ski P,Schuitemaker JH,Hilkens CM,Kapsenberg ML||Journal of immunology (Baltimore, Md. : 1950) (161:2804)||1998|