Sandwich ELISA analysis of human IL-6 was performed using a Human IL-6 Colorimetric ELISA kit (Product # EH2IL6) by loading 50 ul per well of a mouse anti-human IL-6 biotinylated antibody (Product # M621B) at a concentration of 0.035 ug/ml and 50 ul of Human IL-6 Recombinant Protein (Product # SIL6) at 400, 160, 64, 25.6, 10.24, and 0 pg/ml (both in quadruplicate) across a 4 ug/ml mouse anti-human IL-6 (Product # M620) pre-coated plate. The plate was incubated for 2 hours at room temperature, washed, and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at 1:7000 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 stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
|Tested species reactivity||Human|
|Published species reactivity||Dog, Pig, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human IL-6.|
|Storage buffer||PBS with 4% BSA|
|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.
M621B targets IL-6 in ELISA and WB applications and shows reactivity with Human samples.
The M621B immunogen is recombinant human IL-6.
M621B detects IL-6 which has a predicted molecular weight of approximately 22 kDa.
The M621B IL6 antibody (biotinylated conjugate of clone 7IL6) has successfully been paired as the detection antibody in a sandwich ELISA with capture antibody M620 (clone 5IL6). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Biotinylated antibody M621B (clone 7IL6) and antibody M620 (clone 5IL6) have successfully been used in combination with recombinant IL6 protein SIL6 in ELISA applications.
This gene encodes a cytokine that functions in inflammation and the maturation of B cells. The protein is primarily produced at sites of acute and chronic inflammation, where it is secreted into the serum and induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The functioning of this gene is implicated in a wide variety of inflammation-associated disease states, including suspectibility to diabetes mellitus and systemic juvenile rheumatoid arthritis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Inhibitors of p38 suppress cytokine production in rheumatoid arthritis synovial membranes: does variable inhibition of interleukin-6 production limit effectiveness in vivo?
M621B was used in ELISA to investigate the role of p38 kinase in rheumatoid arthritis synovial membranes
|Page TH,Brown A,Timms EM,Foxwell BM,Ray KP||Arthritis and rheumatism (62:3221)||2010|
Th1 responses to beta-amyloid in young humans convert to regulatory IL-10 responses in Down syndrome and Alzheimer's disease.
M621B 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|
Development of an 8-plex Luminex assay to detect swine cytokines for vaccine development: assessment of immunity after porcine reproductive and respiratory syndrome virus (PRRSV) vaccination.
M621B was used in ELISA to develop a multiplex Luminex assay to measure pig cytokines for vaccine development
|Lawson S,Lunney J,Zuckermann F,Osorio F,Nelson E,Welbon C,Clement T,Fang Y,Wong S,Kulas K,Christopher-Hennings J||Vaccine (28:5356)||2010|
A component of polysaccharide peptidoglycan complex on Lactobacillus induced an improvement of murine model of inflammatory bowel disease and colitis-associated cancer.
M621B was used in ELISA to investigate the role of Lactobacillus polysaccharide peptidoglycan complex in chronic intestinal inflammatory disorders
|Matsumoto S,Hara T,Nagaoka M,Mike A,Mitsuyama K,Sako T,Yamamoto M,Kado S,Takada T||Immunology (128:e170)||2009|
Human mast cell activation with virus-associated stimuli leads to the selective chemotaxis of natural killer cells by a CXCL8-dependent mechanism.
M621B was used in ELISA to study the role of human mast cells in the recruitment of natural killer cells
|Burke SM,Issekutz TB,Mohan K,Lee PW,Shmulevitz M,Marshall JS||Blood (111:5467)||2008|
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
M621B was used in ELISA to study the role of mitochondrial pathway in neutrophil survival
|Saffar AS,Alphonse MP,Shan L,Hayglass KT,Simons FE,Gounni AS||Journal of immunology (Baltimore, Md. : 1950) (178:2535)||2007|
Protein microarray platform for the multiplex analysis of biomarkers in human sera.
M621B 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|
Probiotic Lactobacillus-induced improvement in murine chronic inflammatory bowel disease is associated with the down-regulation of pro-inflammatory cytokines in lamina propria mononuclear cells.
M621B was used in ELISA to study the effect of Lactobacillus casei strain Shirota on the production of interleukin 6 in LPS-stimulated murine large intestinal lamina propria mononuclear cells
|Matsumoto S,Hara T,Hori T,Mitsuyama K,Nagaoka M,Tomiyasu N,Suzuki A,Sata M||Clinical and experimental immunology (140:417)||2005|
Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.
M621B 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|
Tumor-infiltrating lymphocyte secretion of IL-6 antagonizes tumor-derived TGF-beta 1 and restores the lymphokine-activated killing activity.
M621B was used in ELISA to investigate the function of interleukin-6 from tumor-infiltrating lymphocytes
|Hsiao YW,Liao KW,Hung SW,Chu RM||Journal of immunology (Baltimore, Md. : 1950) (172:1508)||2004|
Serum cytokine levels, cigarette smoking and airway responsiveness among pregnant women.
M621B was used in ELISA to investigate the relationship between serum cytokine levels, cigarette smoking and airway responsiveness among pregnant women
|Tsunoda M,Litonjua AA,Kuniak MP,Weiss ST,Satoh T,Guevarra L,Tollerud DJ||International archives of allergy and immunology (130:158)||2003|
|Not Applicable||Not Cited||
Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.
M621B was used in ELISA to test if factors released from human alveolar macrophages exposed to air pollution particles accelerate the maturation of granulocyte precursors
|Suwa T,Hogg JC,Vincent R,Mukae H,Fujii T,van Eeden SF||Experimental lung research (28:1)||2002|
Release of vasoactive cytokines by antibody-enhanced dengue virus infection of a human mast cell/basophil line.
M621B was used in ELISA to investigate the role of mast cells/basophils in dengue pathogenesis
|King CA,Marshall JS,Alshurafa H,Anderson R||Journal of virology (74:7146)||2000|
Regulation of interleukin-12 by complement receptor 3 signaling.
M621B was used in ELISA to assess the interferon-alpha production in peripheral blood mononuclear cell
|Marth T,Kelsall BL||The Journal of experimental medicine (185:1987)||1997|
Sustained high levels of circulating chaperoned interleukin-6 after active specific cancer immunotherapy.
M621B was used in ELISA to investigate the changes of interleukin 6 level in patients with cancer immunotherapy
|May LT,Patel K,García D,Ndubuisi MI,Ferrone S,Mittelman A,Mackiewicz A,Sehgal PB||Blood (84:1887)||1994|
Interleukin-6 fused to an anti-idiotype antibody in a vaccine increases the specific humoral immune response against CA125+ (MUC-16) ovarian cancer.
M621B was used in western blot to study the capacity of IL-6 fused to an anti-idiotype antibody in a vaccine to increase the specific humoral immune response against CA125+ ovarian cancer
|Reinartz S,Hombach A,Köhler S,Schlebusch H,Wallwiener D,Abken H,Wagner U||Cancer research (63:3234)||2003|
A nonsteroidal glucocorticoid receptor antagonist.
M621B was used in immunocytochemistry to characterize a potent nonsteroidal antagonist against glucocorticoid receptor
|Miner JN,Tyree C,Hu J,Berger E,Marschke K,Nakane M,Coghlan MJ,Clemm D,Lane B,Rosen J||Molecular endocrinology (Baltimore, Md.) (17:117)||2003|