|Applications||ELISA, IHC, WB|
|Product Name||Anti-IL4 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-4(IL4) detection. Tested with WB, IHC-P, ELISA in Human.|
|Cite This Product||Anti-IL4 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1009)|
|Immunogen||E. coli-derived human IL-4 recombinant protein(Position: H25-S153).|
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-4 using anti- IL-4 antibody (RP1009).
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-4 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-4 antigen affinity purified polyclonal antibody (Catalog # RP1009) 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-4 at approximately 14KD. The expected band size for IL-4 is at 14KD.
Protein Target Info (Source: Uniprot.org)
|Alternative Names||Interleukin-4;IL-4;B-cell stimulatory factor 1;BSF-1;Binetrakin;Lymphocyte stimulatory factor 1;Pitrakinra;IL4;|
|Molecular Weight||17492 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Participates in at least several B-cell activation processes as well as of other cell types. It is a costimulator of DNA-synthesis. It induces the expression of class II MHC molecules on resting B-cells. It enhances both secretion and cell surface expression of IgE and IgG1. It also regulates the expression of the low affinity Fc receptor for IgE (CD23) on both lymphocytes and monocytes.|
|Background||Interleukin-4 (IL-4), also knowns as a B-cell stimulatory factor1 (BSF1), is an immunomodulatory cytokine, which can inhibit the growth of tumour cells.1 The human cDNA contains a single open reading frame encoding a protein of 153 amino acids, including a putative signal peptide. IL-4 may act as an autocrine growth factor in pancreatic cancer cells and also give rise to the possibility that cancer-derived IL-4 may suppress cancer-directed immunosurveillance in vivo in addition to its growth-promoting effects, thereby facilitating pancreatic tumor growth and metastasis.1 The mouse and human genes and their protein products show structural and functional similarities. The human IL-4 gene, which occurs as a single copy in the haploid genome, is mapped on chromosome 5.2 The standard product used in this kit is recombinant human IL-4, consisting of 130 amino acids with the molecular mass of 14KDa.|
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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.,