M620

antibody from Invitrogen Antibodies

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

Provider product page for M620

Western blot

ELISA

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. 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

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