Antibody data
- Antibody Data
- Antigen structure
- References [25]
- Comments [0]
- Validations
- Western blot [1]
- Flow cytometry [3]
- Other assay [4]
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Validation data
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- Product number
- M620 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IL-6 Monoclonal Antibody (5IL6)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- M620 was successfully to detect IL-6 in Flow application. M620 targets IL-6 in ELISA, and Western blot applications and shows reactivity with human samples. The M620 immunogen is recombinant human IL-6. M620 detects IL-6 which has a predicted molecular weight of approximately 22 kDa. The M620 IL6 antibody (clone 5IL6) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M621B (biotinylated conjugate of clone 7IL6). Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection. Antibody M620 (clone 5IL6) and biotinylated antibody M621B (clone 7IL6) have successfully been used in combination with recombinant IL6 protein SIL6 in ELISA applications. This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 5IL6
- Vial size
- 500 µg
- Concentration
- 1.0 mg/mL
- Storage
- -20°C
Submitted references Hepcidin Increases Cytokines in Alzheimer's Disease and Down's Syndrome Dementia: Implication of Impaired Iron Homeostasis in Neuroinflammation.
Impaired Iron Homeostasis and Haematopoiesis Impacts Inflammation in the Ageing Process in Down Syndrome Dementia.
Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury.
Sevoflurane exerts protective effects on liver ischemia/reperfusion injury by regulating NFKB3 expression via miR-9-5p.
Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children.
Interleukin-6 is a potential therapeutic target in interleukin-6 dependent, estrogen receptor-α-positive breast cancer.
Development and validation of panoptic Meso scale discovery assay to quantify total systemic interleukin-6.
Inhibitors of p38 suppress cytokine production in rheumatoid arthritis synovial membranes: does variable inhibition of interleukin-6 production limit effectiveness in vivo?
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.
A component of polysaccharide peptidoglycan complex on Lactobacillus induced an improvement of murine model of inflammatory bowel disease and colitis-associated cancer.
Human mast cell activation with virus-associated stimuli leads to the selective chemotaxis of natural killer cells by a CXCL8-dependent mechanism.
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
Protein microarray platform for the multiplex analysis of biomarkers in human sera.
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.
Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.
A nonsteroidal glucocorticoid receptor antagonist.
Serum cytokine levels, cigarette smoking and airway responsiveness among pregnant women.
Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.
Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.
Release of vasoactive cytokines by antibody-enhanced dengue virus infection of a human mast cell/basophil line.
Release of vasoactive cytokines by antibody-enhanced dengue virus infection of a human mast cell/basophil line.
Regulation of interleukin-12 by complement receptor 3 signaling.
Regulation of interleukin-12 by complement receptor 3 signaling.
Sustained high levels of circulating chaperoned interleukin-6 after active specific cancer immunotherapy.
Raha AA, Ghaffari SD, Henderson J, Chakraborty S, Allinson K, Friedland RP, Holland A, Zaman SH, Mukaetova-Ladinska EB, Raha-Chowdhury R
Frontiers in aging neuroscience 2021;13:653591
Frontiers in aging neuroscience 2021;13:653591
Impaired Iron Homeostasis and Haematopoiesis Impacts Inflammation in the Ageing Process in Down Syndrome Dementia.
Raha-Chowdhury R, Raha AA, Henderson J, Ghaffari SD, Grigorova M, Beresford-Webb J, Allinson K, Chakraborty S, Holland A, Zaman SH
Journal of clinical medicine 2021 Jun 29;10(13)
Journal of clinical medicine 2021 Jun 29;10(13)
Pulsed focused ultrasound enhances the therapeutic effect of mesenchymal stromal cell-derived extracellular vesicles in acute kidney injury.
Ullah M, Liu DD, Rai S, Razavi M, Concepcion W, Thakor AS
Stem cell research & therapy 2020 Sep 14;11(1):398
Stem cell research & therapy 2020 Sep 14;11(1):398
Sevoflurane exerts protective effects on liver ischemia/reperfusion injury by regulating NFKB3 expression via miR-9-5p.
Liao X, Zhou S, Zong J, Wang Z
Experimental and therapeutic medicine 2019 Apr;17(4):2632-2640
Experimental and therapeutic medicine 2019 Apr;17(4):2632-2640
Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein 180L Single-Nucleotide Polymorphism Is Associated With Susceptibility to Recurrent Pneumococcal Lower Respiratory Tract Infections in Children.
Siebert JN, Hamann L, Verolet CM, Gameiro C, Grillet S, Siegrist CA, Posfay-Barbe KM
Frontiers in immunology 2018;9:1780
Frontiers in immunology 2018;9:1780
Interleukin-6 is a potential therapeutic target in interleukin-6 dependent, estrogen receptor-α-positive breast cancer.
Casneuf T, Axel AE, King P, Alvarez JD, Werbeck JL, Verhulst T, Verstraeten K, Hall BM, Sasser AK
Breast cancer (Dove Medical Press) 2016;8:13-27
Breast cancer (Dove Medical Press) 2016;8:13-27
Development and validation of panoptic Meso scale discovery assay to quantify total systemic interleukin-6.
Chaturvedi S, Siegel D, Wagner CL, Park J, van de Velde H, Vermeulen J, Fung MC, Reddy M, Hall B, Sasser K
British journal of clinical pharmacology 2015 Oct;80(4):687-97
British journal of clinical pharmacology 2015 Oct;80(4):687-97
Inhibitors of p38 suppress cytokine production in rheumatoid arthritis synovial membranes: does variable inhibition of interleukin-6 production limit effectiveness in vivo?
Page TH, Brown A, Timms EM, Foxwell BM, Ray KP
Arthritis and rheumatism 2010 Nov;62(11):3221-31
Arthritis and rheumatism 2010 Nov;62(11):3221-31
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.
Lawson S, Lunney J, Zuckermann F, Osorio F, Nelson E, Welbon C, Clement T, Fang Y, Wong S, Kulas K, Christopher-Hennings J
Vaccine 2010 Jul 19;28(32):5356-64
Vaccine 2010 Jul 19;28(32):5356-64
A component of polysaccharide peptidoglycan complex on Lactobacillus induced an improvement of murine model of inflammatory bowel disease and colitis-associated cancer.
Matsumoto S, Hara T, Nagaoka M, Mike A, Mitsuyama K, Sako T, Yamamoto M, Kado S, Takada T
Immunology 2009 Sep;128(1 Suppl):e170-80
Immunology 2009 Sep;128(1 Suppl):e170-80
Human mast cell activation with virus-associated stimuli leads to the selective chemotaxis of natural killer cells by a CXCL8-dependent mechanism.
Burke SM, Issekutz TB, Mohan K, Lee PW, Shmulevitz M, Marshall JS
Blood 2008 Jun 15;111(12):5467-76
Blood 2008 Jun 15;111(12):5467-76
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
Saffar AS, Alphonse MP, Shan L, Hayglass KT, Simons FE, Gounni AS
Journal of immunology (Baltimore, Md. : 1950) 2007 Feb 15;178(4):2535-41
Journal of immunology (Baltimore, Md. : 1950) 2007 Feb 15;178(4):2535-41
IgE modulates neutrophil survival in asthma: role of mitochondrial pathway.
Saffar AS, Alphonse MP, Shan L, Hayglass KT, Simons FE, Gounni AS
Journal of immunology (Baltimore, Md. : 1950) 2007 Feb 15;178(4):2535-41
Journal of immunology (Baltimore, Md. : 1950) 2007 Feb 15;178(4):2535-41
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 2006 Oct 20;316(1-2):1-7
Journal of immunological methods 2006 Oct 20;316(1-2):1-7
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.
Matsumoto S, Hara T, Hori T, Mitsuyama K, Nagaoka M, Tomiyasu N, Suzuki A, Sata M
Clinical and experimental immunology 2005 Jun;140(3):417-26
Clinical and experimental immunology 2005 Jun;140(3):417-26
Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.
Schotte H, Schlüter B, Willeke P, Mickholz E, Schorat MA, Domschke W, Gaubitz M
Rheumatology (Oxford, England) 2004 Aug;43(8):960-4
Rheumatology (Oxford, England) 2004 Aug;43(8):960-4
A nonsteroidal glucocorticoid receptor antagonist.
Miner JN, Tyree C, Hu J, Berger E, Marschke K, Nakane M, Coghlan MJ, Clemm D, Lane B, Rosen J
Molecular endocrinology (Baltimore, Md.) 2003 Jan;17(1):117-27
Molecular endocrinology (Baltimore, Md.) 2003 Jan;17(1):117-27
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 2003 Feb;130(2):158-64
International archives of allergy and immunology 2003 Feb;130(2):158-64
Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.
Suwa T, Hogg JC, Vincent R, Mukae H, Fujii T, van Eeden SF
Experimental lung research 2002 Jan-Feb;28(1):1-18
Experimental lung research 2002 Jan-Feb;28(1):1-18
Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.
Suwa T, Hogg JC, Vincent R, Mukae H, Fujii T, van Eeden SF
Experimental lung research 2002 Jan-Feb;28(1):1-18
Experimental lung research 2002 Jan-Feb;28(1):1-18
Release of vasoactive cytokines by antibody-enhanced dengue virus infection of a human mast cell/basophil line.
King CA, Marshall JS, Alshurafa H, Anderson R
Journal of virology 2000 Aug;74(15):7146-50
Journal of virology 2000 Aug;74(15):7146-50
Release of vasoactive cytokines by antibody-enhanced dengue virus infection of a human mast cell/basophil line.
King CA, Marshall JS, Alshurafa H, Anderson R
Journal of virology 2000 Aug;74(15):7146-50
Journal of virology 2000 Aug;74(15):7146-50
Regulation of interleukin-12 by complement receptor 3 signaling.
Marth T, Kelsall BL
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
Regulation of interleukin-12 by complement receptor 3 signaling.
Marth T, Kelsall BL
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
Sustained high levels of circulating chaperoned interleukin-6 after active specific cancer immunotherapy.
May LT, Patel K, García D, Ndubuisi MI, Ferrone S, Mittelman A, Mackiewicz A, Sehgal PB
Blood 1994 Sep 15;84(6):1887-95
Blood 1994 Sep 15;84(6):1887-95
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IL-6 Monoclonal Antibody (5IL6) (Product # M620) and a 24 kDa band corresponding to IL-6 was observed across to be upregulated upon treatment of HUVEC cells with LPS and Brefeldin A. Whole cell extracts (50 µg lysate) of HUVEC (Lane 1) and HUVEC treated with LPS and Brefeldin A (0.5 µg/mL of LPS for 24 h and 300 ng/mL of Brefeldin A for the last 20 h of LPS stimulation) (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:50,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Expression of IL-6 was observed to be to be upregulated upon treatment of HUVEC cells with LPS and Brefeldin A.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of IL-6 on LPS treated THP-1 cells. Cells were fixed with 4% formaldehyde for 30 min on ince and permeabilized with IC permeabilization buffer (Product # PB001). After incubation with blocking buffer (Product # 37525) for 30 min on ice, cells were then stained with anti-IL-6 mouse monoclonal antibody (Product # M620) at indicated dilution with IC permeabilization buffer for 30 min on ice. After washing with ice-cold IC permeabilization buffer for 3 times, the cells were stained with DyLight 488 goat anti-mouse secondary antibody (Product # 35502) for 30 min on ice. A representative 10,000 cells were acquired for each sample.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of IL-6 on LPS treated THP-1 cells. Cells were fixed with 4% formaldehyde for 30 min on ince and permeabilized with IC permeabilization buffer (Product # PB001). After incubation with blocking buffer (Product # 37525) for 30 min on ice, cells were then stained with anti-IL-6 mouse monoclonal antibody (Product # M620) at 1:100 dilution with IC permeabilization buffer for 30 min on ice. After washing with ice-cold IC permeabilization buffer for 3 times, the cells were stained with DyLight 488 goat anti-mouse secondary antibody (Product # 35502) for 30 min on ice. A representative 10,000 cells were acquired for each sample.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of IL-6 on PHA-M stimulated THP-1 cells with or without LPS treatment. Cells were fixed with 4% formaldehyde for 30 min on ince and permeabilized with IC permeabilization buffer (Product # PB001). After incubation with blocking buffer (Product # 37525) for 30 min on ice, cells were then stained with anti-IL-6 mouse monoclonal antibody (Product # M620) at 1:100 dilution with IC permeabilization buffer for 30 min on ice. After washing with ice-cold IC permeabilization buffer for 3 times, the cells were stained with DyLight 488 goat anti-mouse secondary antibody (Product # 35502) for 30 min on ice. A representative 10,000 cells were acquired for each sample.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Sandwich ELISA analysis of human IL-6 was performed using a Human IL-6 Colorimetric ELISA kit (Product # EH2IL6) by loading 50 µL per well of a mouse anti-human IL-6 biotinylated antibody (Product # M621B) at a concentration of 0.035 µg/mL and 50 µL 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 µg/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 µL 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 4 Inflammatory cytokines. a Immunohistochemistry staining for inflammatory markers TNF-alpha and IL-6 in kidney tissue. b Western blot on kidney tissue measuring inflammatory markers TNF-alpha and NF-kappaB (left), alongside their quantification (right). c Quantitative real-time PCR on kidney tissue measuring inflammatory markers Nfkb , Il6 , and Tnfa . d ELISA measurement of blood serum concentrations of cytokines IL-1beta, IL-6, and TNF-alpha. Each group has n = 5 mice. Significant difference a p < 0.05: relative to untreated control; b p < 0.05: relative to AKI; c p < 0.05: relative to AKI-EV; d p < 0.05: relative to AKI-EV-pFUS
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 IL-6 could be involved in host defense in AD and DS brain. ELISA based analysis of IL-6 in serum samples showing increased levels in AD and highest level in DS when compared with hepcidin ( p < 0.0001; A ). Samples were analyzed from AD and age matched older controls (OC), IL-6 levels were significantly higher in AD serum ( B ; p < 0.0001). Ferritin (FTH) was present close to a SP in DS brain analyzed by double immunofluorescence, and did not co-localize with Abeta42 positive plaques (C) . Another control brain section from white matter tract (WMT) of corpus callosum (CC), labeled with hepcidin, expression being visible in the WMT (in the oligodendrocytes), whereas IL-1beta being present in the microglia with minimal co-localization (D-F) . Brain sections from AD (HP, hippocampus) and DS (SFG) when stained with hepcidin and IL-1beta, revealed similarly the presence of hepcidin (punctuate, vesicular appearance) in the hippocampal neurons, whereas IL-1beta was visible only in the microglia with minimum co-localization (G,H) . When control and DS brain sections of HP and SFG were stained for hepcidin and IL-6, there was less IL-6 visible in the control brain (I) , whereas there was a higher co-localization seen in the granule cells of hippocampus of DS subjects (J-L) . Scale bar: (C and G) = 20 mum, (D-F) = 50 mum, (H-L) = 25 mum. *** p < 0.001, **** p < 0.0001.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 Inflammatory changes in the DS brain were identified by TREM2, IL-1beta and IL-6. To investigate the inflammatory changes in DS, the brain sections from DS and control subjects were stained with microglia marker (Iba1) and TREM2. In control brains, Iba1-positive ramified microglia were visible in the cortex, whereas TREM2 was noticed in the neurons close to the blood vessels ( A ). In the DS brain sections, activated microglia was visible around the SP and limited co-localisation with TREM2 in the microglia ( B ). For sections from the HP and DG of DS brain when stained with Iba1 and TREM2, Iba1-positive activated microglia were present close to the neurons, whereas TREM2 expression was visible only in the damaged DG granule cells with limited co-localisation ( C , D ). The white matter (WM) close to the blood vessels (BV) in the DS brain showed large numbers of Iba1-positive microglia, close to the blood vessels, that might be involved in the clearing process of damaged oligodendrocytes, and TREM2-positive macrophages might be entering from blood vessels ( E ). For another DS brain section, close to the BV, when stained with hepcidin and IL-6, the staining patterns suggested that both proteins might be entering from the blood vessels ( F ). The brain sections from controls and DS subjects (from cortex) were stained with TREM2 and pro-inflammatory marker IL-1beta. In control, TREM2 was visible in the neurons ( G ) and in other glial cells, and only very faint IL-1bet