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 biotin conjugated IL-4 rat monoclonal antibody (Product # M450B) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a pre-diluted High Sensitivity Streptavidin-HRP antibody (Product # 21134) at a dilution of 1:50 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|
|Published species reactivity||Human|
|Host / Isotype||Rat / IgG1|
|Immunogen||Recombinant human IL-4 (CHO cell-derived)|
|Storage buffer||PBS with 4% BSA|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.003-12.5 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.
M450B targets IL-4 in WB and ELISA applications and shows reactivity with Human samples.
The M450B immunogen is recombinant human IL-4 (CHO cell-derived).
M450B detects IL-4 which has a predicted molecular weight of approximately 15 kDa.
This antibody is produced by injecting Rat IgG secreting hybridoma cells into the peritoneum of mice. The resulting ascites is collected from the mice and the antibody is purified.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Interleukin-13 mucosal production in Helicobacter pylori-related gastric diseases.
M450B was used in ELISA to study the mucosal production of interleukin 13 in Helicobacter pylori-related gastric diseases
|Marotti B,Rocco A,De Colibus P,Compare D,de Nucci G,Staibano S,Tatangelo F,Romano M,Nardone G||Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver (40:240)||2008|
Antigen-independent Th2 cell differentiation by stimulation of CD28: regulation via IL-4 gene expression and mitogen-activated protein kinase activation.
M450B was used in ELISA to investigate the role of interleukin 4 gene expression
|Skapenko A,Lipsky PE,Kraetsch HG,Kalden JR,Schulze-Koops H||Journal of immunology (Baltimore, Md. : 1950) (166:4283)||2001|
Over-expression of interleukin 10 in mucosal T cells of patients with active ulcerative colitis.
M450B was used in immunohistochemistry to study the involvement of interleukin 10 in active ulcerative colitis
|Melgar S,Yeung MM,Bas A,Forsberg G,Suhr O,Oberg A,Hammarstrom S,Danielsson A,Hammarstrom ML||Clinical and experimental immunology (134:127)||2003|