|Applications||ELISA, IHC, WB|
|Product Name||Anti-IL10 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-10(IL10) detection. Tested with WB, IHC-P, ELISA in Human.|
|Cite This Product||Anti-IL10 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1014)|
|Immunogen||E. coli-derived human IL-10 recombinant protein(Position: S19-N178).|
Assay Dilutions Overview
ELISA , 0.1-0.5μg/ml, Human, -
Western blot, 0.1-0.5μg/ml, Human
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, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure. Western blot analysis of IL-10 using anti- IL-10 antibody (RP1014).
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-10 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-10 antigen affinity purified polyclonal antibody (Catalog # RP1014) 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-10 at approximately 19KD. The expected band size for IL-10 is at 19KD.
Protein Target Info (Source: Uniprot.org)
|Tissue Specificity||Produced by a variety of cell lines, including T-cells, macrophages, mast cells and other cell types.|
|Alternative Names||Interleukin-10;IL-10;Cytokine synthesis inhibitory factor;CSIF;IL10;|
|Molecular Weight||20517 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Inhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells.|
|Background||Interleukin-10(IL-10 or IL10), also known as human cytokine synthesis inhibitory factor(CSIF), is an anti-inflammatory cytokine. In humans IL-10 is encoded by the IL10 gene. It is capable of inhibiting synthesis of pro-inflammatory cytokines like IFN-gamma, IL-2, IL-3, TNFalpha and GM-CSF made by cells such as macrophages and regulatory T-cells.IL-10 also displays potent abilities to suppress the antigen presentation capacity of antigen presenting cells. Kim et al.(1992) showed that the mouse IL 10 gene contains 5 exons and spans about 5.2 kb of genomic DNA. Eskdale et al.(1997) mapped the IL10 gene to the junction between 1q31 and 1q32.|
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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.,