IL6 antibody

• Catalog Number: orb750681
• Category: Antibodies
• Description: IL6 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: DOT, ELISA, IHC, IP, WB
• Reactivity: Human
• Isotype: Antiserum
• Immunogen: This whole rabbit serum was prepared by repeated immunizations with recombinant human IL-6 produced in E.coli.
• Concentration: 80 mg/mL
• Dilution range: ELISA: 1:1,000 - 1:5,000, IHC: 1:400 - 1:800, IP: 1:400 - 1:800, WB: 1:500 - 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-IL-6 antiserum detects recombinant and native IL-6 present in body fluids and cell supernatants in various assays (ie. IL-1 stimulated IL-6 production from fibroblasts). This product has minimal reactivity with mouse IL-6.
• Conjugation: Unconjugated
• UniProt ID: P05231
• NCBI: NP_000591.1
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: None
• Alternative names: HSF, Hybridoma growth factor, HGF, Hybridoma plasm
• Note: For research use only
• Application notes: IL-6 antibody has been tested in western blot. This antibody is suitable for use in neutralizations, ELISA, radioimmunoassay, and immunoprecipitation. Reactivity in other immunoassays is unknown. Expect ~23.7kDa. In Western blot analysis of natural cell products or human body fluids, multiple bands of IL-6 will appear due to the variable amount of glycosylation on the molecule. For immunoblot use the supernatants or lysates of 2 x 106 endotoxin-stimulated human peripheral blood mononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1% (v/v) human serum plus 10ng/mL E. coli LPS. For immunoprecipitation, pre-clearing with a non-specific rabbit IgG is helpful to reduce background. The antiserum is useful for neutralization of human of IL-6 activity in bioassays. For neutralization, incubate the sample with a 1:400 dilution of the antiserum for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG (heat inactivated) is recommended. In neutralization experiments in vitro, this antibody does not result in enhanced activity of IL-6. However, because antibodies to IL-6 may act as a soluble receptor in vivo, some antibodies to IL-6 act as carriers and enhance IL-6 activity.
• Expiration Date: 12 months from date of receipt.

Western Blot of Rabbit Anti-IL-6 Antibody. Lane 1: Opal Prestained Molecular Weight Marker. Lane 2: Human Rec. IL-6/HeLa Whole Cell Lysate [0.05 µg/10 µg]. Lane 3: Human Rec. IL-6/HeLa Whole Cell Lysate [0.02 µg/10 µg]. Lane 4: Human Rec. IL-6/HeLa Whole Cell Lysate [0.01 µg/10 µg]. Lane 5: HeLa Whole Cell Lysate (p/n orb348668) [10 µg]. Primary Antibody: Anti-IL-6 at 1:1000 overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP conjugate (p/n orb347654) at 1:70000 for 30 mins at RT. Block: BlockOut Buffer (p/n orb348644). Predicted MW: ~23 kDa. Observed MW: ~19 Exposure: 2 seconds.

Western blot using Biorbyt's Anti-IL6 antibody. Protein was resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. The blot shows detection of a band ~21 kDa in size corresponding to anti-IL6 antibody. Molecular weight markers are also shown (MW). After transfer, the membrane was blocked for 30 minutes with 1% BSA-TBST. Detection occurred using peroxidase conjugated Goat anti-Rabbit IgG (p/n orb347654) secondary antibody diluted 1:40000 in blocking buffer (p/n orb348637) for 30 min at RT followed by reaction with FemtoMax™ chemiluminescent substrate.