M620

antibody from Invitrogen Antibodies

Targeting: IL6 BSF2, HGF, HSF, IFNB2, IL-6

Provider product page for M620

Western blot

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [25]
Comments [0]
Validations
Western blot [1]
Flow cytometry [3]
Other assay [4]

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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.
Antibody clone number: 5IL6
Concentration: 1.0 mg/mL

IL6 protein structure - M620 shown in red.

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.

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 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: 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.

Supportive validation

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

Supportive validation

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.

Supportive validation

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