|Applications||ELISA, IHC, WB|
|Product Name||Anti-IL3 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-3(IL3) detection. Tested with WB, IHC-P, ELISA in Human.|
|Cite This Product||Anti-IL3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1008)|
|Immunogen||E. coli-derived human IL-3 recombinant protein(Position: A20-F152).|
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-3 using anti- IL-3 antibody (RP1008).
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-3 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-3 antigen affinity purified polyclonal antibody (Catalog # RP1008) 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-3 at approximately 15KD. The expected band size for IL-3 is at 15KD.
Protein Target Info (Source: Uniprot.org)
|Tissue Specificity||Activated T-cells, mast cells, natural killer cells.|
|Alternative Names||Interleukin-3;IL-3;Hematopoietic growth factor;Mast cell growth factor;MCGF;Multipotential colony-stimulating factor;P-cell-stimulating factor;IL3;|
|Molecular Weight||17233 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Granulocyte/macrophage colony-stimulating factors are cytokines that act in hematopoiesis by controlling the production, differentiation, and function of 2 related white cell populations of the blood, the granulocytes and the monocytes-macrophages.|
|Background||The gene IL-3 encodes interleukin 3, a hematopoietic colony-stimulating factor(CSF) that is capable of supporting the proliferation of a broad range of hematopoietic cell types.1 Interleukin-3(IL-3), a protein of 140 amino acids, is chemically synthesized by means of an automated peptide synthesizer and is shown to have the biological activities attributed to native IL-3.2 The cDNA sequence for murine interleukin-3, one of the colony stimulating factors that regulate haematopoiesis, codes for a polypeptide of 166 amino acids including a putative signal peptide.3 The mouse IL 3 gene is located on chromosome 11.4 The human gene encoding IL 3 is tandemly arrayed on the long arm of chromosome 5.5 The standard product used in this kit is recombinant human IL-3, consisting of 133 amino acids with the molecular mass of 15KDa.|
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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.,