Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Immunocytochemistry [1]
- Flow cytometry [3]
- Other assay [3]
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Validation data
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- Product number
- 701028 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IL-6 Recombinant Rabbit Monoclonal Antibody (4H16L21)
- Antibody type
- Monoclonal
- Antigen
- Other
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 4H16L21
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Effect of PM2.5 on invasion and proliferation of HeLa cells and the expression of inflammatory cytokines IL-1 and IL-6.
Four assay designs and on-chip calibration: gadgets for a sepsis protein array.
Huang K, Li W, Chen Y, Zhu J
Oncology letters 2018 Dec;16(6):7068-7073
Oncology letters 2018 Dec;16(6):7068-7073
Four assay designs and on-chip calibration: gadgets for a sepsis protein array.
Buchegger P, Preininger C
Analytical chemistry 2014 Mar 18;86(6):3174-80
Analytical chemistry 2014 Mar 18;86(6):3174-80
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of IL-6 was performed using 70% confluent log phase HUVEC cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with IL-6 Recombinant Rabbit Monoclonal Antibody (4H16L21) (Product # 701028) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly Golgi Complex and Endoplasmic Reticulum localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
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 # 701028) or IgG control 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 rabbit monoclonal antibody (Product # 701028) 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-rabbit secondary antibody (Product # 35552) 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 rabbit monoclonal antibody (Product # 701028) 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-rabbit secondary antibody (Product # 35552) 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
- Figure 5. ELISA. ELISA was used to detect the expression of IL-1 and IL-6 in two groups of cells. The OD values of IL-1 and IL-6 in HeLa cells treated with 10 ug/ml PM2.5 were significantly higher than those in NC group. *P
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Indirect ELISA was performed using various dilutions of Anti-IL-6 Rabbit Monoclonal Antibody (Product # 701028) to detect IL-6 recombinant protein coated onto the plate. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody was determined.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3. Western blotting. Compared with HeLa cells in NC group, the relative expression levels of (A) IL-1 and (B) IL-6 proteins in HeLa cells treated with 10 ug/ml PM2.5 were significantly increased. (C) Western blots of IL-1 and IL-6 protein in the two groups of cells are shown. *P