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 # M2IL4I) 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 / IgG1|
|Immunogen||Recombinant human IL-4 purified from Pichia pastoris|
|Storage buffer||PBS with 5% trehalose|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.001-20 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.
M2IL4I targets IL-4 in ELISA and Western blot applications and shows reactivity with Human samples.
The M2IL4I immunogen is recombinant human IL-4 purified from Pichia pastoris.
M2IL4I detects IL-4 which has a predicted molecular weight of approximately 15 kDa.
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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