Product nameAnti-IL-6 antibody
See all IL-6 primary antibodies
DescriptionMouse monoclonal to IL-6
Tested applicationsSuitable for: IHC-Fr, ICC/IF, Neutralising, Sandwich ELISA, IHC-P, WBmore details
Species reactivityReacts with: Rat, Human
- Purchase matching WB positive control:Recombinant Human IL-6 protein
- WB: Human spleen and lung tissue lysates, Rat spleen and lung tissue lysates, Recombinant human IL6 protein (ab9627); IHC-P: Human spleen tissue, human cervical squamous cell carcinoma tissue; IHC-Fr: Murine neural tissue; ICC/IF: Rat primary microglial cells.
FormLyophilised:Reconstitute with 500µl of sterile water. Please note that if you receive this product in liquid form it has already been reconstituted as described and no further reconstitution is necessary.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Concentration information loading...
PurityProtein A purified
ELISA pair antibody
Our Abpromise guarantee covers the use of ab9324 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|Neutralising||Use at an assay dependent concentration. To yield one-half maximal inhibition [ND50] of the biological activity of hIL-6 (0.5 ng/ml), a concentration of 19.0-23.0 ug/ml of this antibody is required.|
|Sandwich ELISA||Use at an assay dependent concentration.
In a sandwich ELISA (assuming 100 µl/well), a concentration of 1.0-2.0 µg/mL of this antibody will detect at least 100 pg/mL of recombinant human IL-6 when used with Rabbit polyclonal to IL6 (Biotin) (ab84251) as the detection antibody at a concentration of approximately 0.10-0.20 µg/mL.
|IHC-P||Use a concentration of 0.125 µg/ml.|
|WB||Use at an assay dependent concentration.
To detect human IL-6 by Western blot analysis this antibody can be used at a concentration of 0.50-1.0 μg/mL. When used in conjunction with compatible secondary reagents the detection limit for recombinant human IL-6 is 1.0-2.0 ng/lane, under reducing conditions and non-reducing conditions.
FunctionCytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoeitic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance.
Involvement in diseaseGenetic variations in IL6 are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis.
Note=A IL6 promoter polymorphism is associated with a lifetime risk of development of Kaposi sarcoma in HIV-infected men.
Sequence similaritiesBelongs to the IL-6 superfamily.
modificationsN- and O-glycosylated.
- Information by UniProt
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All lanes : Anti-IL-6 antibody (ab9324) at 1/2000 dilution
Lane 1 : Human Spleen
Lane 2 : Human Lung
Lane 3 : Rat Spleen
Lane 4 : Rat Lung
Lysates/proteins at 20 µg per lane.
All lanes : 800CW Goat Anti-Mouse IgG at 1/10000 dilution
Performed under reducing conditions.
Observed band size: 17,25 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible multimer)
This antibody was raised against an immunogen that is predicted to recognize the glycosylated form of IL6 as well as the IL6delta4 splice variant. The predicted molecular weights are 25 kDa and 17 kDa respectively. The band observed at 50 kDa may represent multimers of IL6 as reported in the literature.
IHC image of IL6 staining in human spleen formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9324, 1in250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Immunohistochemical analysis of PFA-fixed frozen murine neural tissue sections, labelling IL6 with ab9324 at a dilution of 1/1000 incubated for 24 hours at 4°C in 0.1M PBST with 10% donkey serum. Permeabilization was with 0.1M PBS and 3% Triton X. Blocking was with 10% serum incubated for 1 hour at 24°C. Secondary was a mouse monoclonal Alexa Fluor® 568 conjugate at 1/1000.
ab9324 staining IL6 in human cervical squamous cell carcinoma tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections). Tissue underwent heat mediated antigen retrieval in sodium citrate buffer (pH 6.0). The primary antibody was used at 0.125 ug/ml and incubated with sample at 4°C overnight. A HRP-labeled polymer detection system was used with a DAB chromogen.
ICC/IF image of IL6 staining (ab9324) in rat primary microglial cell culture. The sections were incubated in 10% serum to block non-specific protein-protein interactions and in 0.3% triton X in 0.1% PBS for 1h to permeabilise the cells. The sections were then incubated with ab9324 (1:1000) overnight at +4°C, followed by Alexa 568 conjugated secondary antibody. Red staining of the cytosol was observed.
This product has been referenced in:
- Jitmana R et al. Role of cardiac mast cells in exercise training-mediated cardiac remodeling in angiotensin II-infused ovariectomized rats. Life Sci 219:209-218 (2019). Read more (PubMed: 30658099) »
- Zhang G et al. Resveratrol alleviates lipopolysaccharide-induced inflammation in PC-12 cells and in rat model. BMC Biotechnol 19:10 (2019). Read more (PubMed: 30691440) »