Western blot analysis of IL-4 was performed by loading 1ug of human recombiannt protein IL-4 (1ug/lane, Product #RIL4I) in non-reducing sample buffer and 8ul PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product #WT4202BOX). Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. IL-4 was detected using a IL-4 mouse monoclonal antibody (Product # M3IL4I) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a HRP conjugated secondary antibody (Product # 62-6520) at a dilution of 1:5000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura Extended Duration Substrate (Product # 34076) and the myECL Imager (Product # 62236).
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Recombinant human IL-4 purified from Pichia pastoris|
|Storage buffer||PBS with 5% trehalose|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.005-20 µg/mL|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
M3IL4I targets IL-4 in ELISA and Western blot applications and shows reactivity with Human samples.
The M3IL4I immunogen is recombinant human IL-4 purified from Pichia pastoris.
M3IL4I detects IL-4 which has a predicted molecular weight of approximately 15 kDa.
The M3IL4I IL4 antibody (clone E10023) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M2IL4BI (biotinylated conjugate of clone E10022). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody M3IL4I (clone E10022) and biotinylated antibody M2IL4BI (clone E10023) have successfully been used in combination with recombinant IL4 protein SIL4 in ELISA applications.
The protein encoded by this gene is a pleiotropic cytokine produced by activated T cells. This cytokine is a ligand for interleukin 4 receptor. The interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. STAT6, a signal transducer and activator of transcription, has been shown to play a central role in mediating the immune regulatory signal of this cytokine. This gene, IL3, IL5, IL13, and CSF2 form a cytokine gene cluster on chromosome 5q, with this gene particularly close to IL13. This gene, IL13 and IL5 are found to be regulated coordinately by several long-range regulatory elements in an over 120 kilobase range on the chromosome. Two alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported.
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