|Product Name||Anti-IL6 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Interleukin-6(IL6) detection. Tested with WB in Human.|
|Cite This Product||Anti-IL6 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1012)|
|Immunogen||E. coli-derived human IL-6 recombinant protein(Position: P29-M212).|
Assay Dilutions Overview
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
Images And Assay Conditions
Figure. Western blot analysis of IL-6 using anti- IL-6 antibody (RP1012).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane : Recombinant Human IL-6 Protein 0.5ng
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- IL-6 antigen affinity purified polyclonal antibody (Catalog # RP1012) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for IL-6 at approximately 20KD. The expected band size for IL-6 is at 20KD.
Protein Target Info (Source: Uniprot.org)
|Alternative Names||Interleukin-6;IL-6;B-cell stimulatory factor 2;BSF-2;CTL differentiation factor;CDF;Hybridoma growth factor;Interferon beta-2;IFN-beta-2;IL6;IFNB2;|
|Molecular Weight||23718 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Cytokine 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. Acts on B-cells, T-cells, hepatocytes, hematopoietic progenitor cells and cells of the CNS. Required for the generation of T(H)17 cells. 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. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation.|
|Background||Interleukin-6(IL-6) is a protein that in humans is encoded by the IL6 gene. IL-6 is an interleukin that acts as both a pro-inflammatory and anti-inflammatory cytokine. It is secreted by T cells and macrophages to stimulate immune response to trauma, especially burns or other tissue damage leading to inflammation. IL-6 is one of the most important mediators of fever and of the acute phase response. IL-6 is also essential for hybridoma growth and is found in many supplemental cloning media such as briclone. Bowcock et al.(1988) assigned the IL6 gene to chromosome 7p21. By in situ hybridization and Southern blot analysis of mouse-human hybrid cell lines, Sutherland et al.(1988) mapped the IL-6 gene to chromosome 7p15.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,