Sandwich ELISA analysis of human IL-1b was performed using a Human IL-1b Colorimetric ELISA Kit (Product # EH2IL1B) by loading 50 ul per well of Human IL-1b Recombinant Protein (Product # SIL1B) in quadruplicate at 400, 160, 64, 25.6, 10.24 and 0 pg/ml across a 5 ug/ml mouse anti-human IL-1b (Product # M421B) pre-coated plate and incubating for 3 hours at room temperature. The plate was washed, then incubated with 50 ul per well of a biotinylated-mouse anti-human IL-1b (Product # M420BB) in quadruplicate at 0.07 ug/ml for 3 hours at room temperature. The plate was washed and incubated with 100 ul per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at 1:3000 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 450-550 nm.
|Tested species reactivity||Human|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Recombinant human IL-1 beta|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||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.
M421B targets IL-1 beta in ELISA, and WB applications and shows reactivity with Human samples.
A precipitate may be present in the solution. Mix for at least five seconds prior to use.
M421B detects IL-1 beta which has a predicted molecular weight of approximately 31 kDa.
The M421B IL1b antibody (clone ILB1-H67) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M420BB (biotinylated conjugate of clone ILB1-H6). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody M421B (clone ILB1-H67) and biotinylated antibody M420BB (clone ILB1-H6) have successfully been used in combination with recombinant IL1 beta protein SIL1B in ELISA applications.
The protein encoded by this gene is a member of the interleukin 1 cytokine family. This cytokine is produced by activated macrophages as a proprotein, which is proteolytically processed to its active form by caspase 1 (CASP1/ICE). This cytokine is an important mediator of the inflammatory response, and is involved in a variety of cellular activities, including cell proliferation, differentiation, and apoptosis. The induction of cyclooxygenase-2 (PTGS2/COX2) by this cytokine in the central nervous system (CNS) is found to contribute to inflammatory pain hypersensitivity. This gene and eight other interleukin 1 family genes form a cytokine gene cluster on chromosome 2.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Th1 responses to beta-amyloid in young humans convert to regulatory IL-10 responses in Down syndrome and Alzheimer's disease.
M421B was used in ELISA to study the role of Th1 responses to beta amyloid in Down syndrome and Alzheimer disease
|Loewenbrueck KF,Tigno-Aranjuez JT,Boehm BO,Lehmann PV,Tary-Lehmann M||Neurobiology of aging (31:1732)||2010|
Protein microarray platform for the multiplex analysis of biomarkers in human sera.
M421B was used in ELISA to develop a protein microarray platform for the multiplex analysis of biomarkers in human sera
|Urbanowska T,Mangialaio S,Zickler C,Cheevapruk S,Hasler P,Regenass S,Legay F||Journal of immunological methods (316:1)||2006|
Relationship between antipyretic effects and cytokine levels in uncomplicated falciparum malaria during different treatment regimes.
M421B was used in ELISA to investigate the effects of antipyretics on cytokine levels in uncomplicated falciparum malaria
|Hugosson E,Montgomery SM,Premji Z,Troye-Blomberg M,Björkman A||Acta tropica (99:75)||2006|
Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.
M421B was used in ELISA to study the effect of etanercept treatment on proinflammatory cytokine-secreting peripheral blood mononuclear cells in rheumatoid arthritic patients
|Schotte H,Schlüter B,Willeke P,Mickholz E,Schorat MA,Domschke W,Gaubitz M||Rheumatology (Oxford, England) (43:960)||2004|
HIV-1 coat protein gp120 stimulates interleukin-1beta secretion from human neuroblastoma cells: evidence for a role in the mechanism of cell death.
M421B was used in ELISA to investigate the role of interleukin 1 beta during the cell death induced by HIV infection
|Corasaniti MT,Bilotta A,Strongoli MC,Navarra M,Bagetta G,Di Renzo G||British journal of pharmacology (134:1344)||2001|
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.
M421B 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|
IL-12-deficient dendritic cells, generated in the presence of prostaglandin E2, promote type 2 cytokine production in maturing human naive T helper cells.
M421B was used in ELISA to investigate the importance of PGE2 during dendritic cell development
|Kali¿ski P,Hilkens CM,Snijders A,Snijdewint FG,Kapsenberg ML||Journal of immunology (Baltimore, Md. : 1950) (159:28)||1997|
Regulation of interleukin-12 by complement receptor 3 signaling.
M421B was used in ELISA to assess the effect of complement receptor 3 on interleukin 12 production
|Marth T,Kelsall BL||The Journal of experimental medicine (185:1987)||1997|
|Not Applicable||Not Cited||
Immune phenotype and intracellular cytokine production of peripheral blood mononuclear cells from postmenopausal patients with osteoporotic fractures.
M421B was used in immunocytochemistry to analyze peripheral blood mononuclear cell surface markers and intracellular cytokine production in postmenopausal osteoporosis patients
|Pietschmann P,Grisar J,Thien R,Willheim M,Kerschan-Schindl K,Preisinger E,Peterlik M||Experimental gerontology (36:1749)||2001|
Intrahepatic expression of interleukin-1beta and tumor necrosis factor-alpha in chronic hepatitis C.
M421B was used in immunohistochemistry to investigate the intrahepatic expression of interleukin 1 beta and tumor necrosis factor alpha in liver specimens with chronic hepatitis C
|Dumoulin FL,Leifeld L,Honecker U,Sauerbruch T,Spengler U||The Journal of infectious diseases (180:1704)||1999|