Sandwich ELISA analysis of human IL-5 was performed by coating wells of a 96-well plate with 110ul of a human IL-5 monoclonal antibody (Product # M551) in duplicate at 1, 2, 4, 5, 7, and 9ug/ml for at least 12 hours. The plate was blocked with a proprietary blocking solution for 1-4 hours at room temperature, and 50ul of mouse IL-5 recombinant protein (Product # SMIL5) was added in replicates of twelve at 400, 160, 64, 25.6, 10.24, and 0pg/ml for 1 hour at room temperature. The plate was washed and incubated 50ul of a biotinylated IL-5 monoclonal antibody (Product # M550B) was added to all test wells at 0.5ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of Streptavidin-HRP (Product # N504) at a 1:10,000 dilution for 30 minutes at room temperature. Detection was performed with 100ul per well of Ultra TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rat / IgG2a|
|Immunogen||Recombinant human IL-6|
|Purification||Caprylic acid precipitation|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
|ELISA (ELISA)||See 2 publications below|
M551 targets IL-5 in ELISA, and WB applications and shows reactivity with Human samples.
The M551 immunogen is recombinant human IL-6.
M551 detects IL-5 which has a predicted molecular weight of approximately 13 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 M551 IL5 antibody (clone 5A10) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M550B (biotinylated conjugate of clone 39D10). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody M551 (clone 5A10) and biotinylated antibody M550B (clone 39D10) have successfully been used in combination with recombinant IL5 protein SIL5 in ELISA applications.
The protein encoded by this gene is a cytokine that acts as a growth and differentiation factor for both B cells and eosinophils. This cytokine is a main regulator of eosinopoiesis, eosinophil maturation and activation. The elevated production of this cytokine is reported to be related to asthma or hypereosinophilic syndromes. The receptor of this cytokine is a heterodimer, whose beta subunit is shared with the receptors for interleukine 3 (IL3) and colony stimulating factor 2 (CSF2/GM-CSF). This gene, together with those for interleukin 4 (IL4), interleukin 13 (IL13), and CSF2, form a cytokine gene cluster on chromosome 5. This cytokine, IL4, and IL13 are found to be regulated coordinately by long-range regulatory elements spread over 120 kilobases on chromosome 5q31.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Natural regulatory T cells: number and function are normal in the majority of patients with lupus nephritis.
M551 was used in ELISA to examine the frequency, phenotype, and function of T regulatory cells in patients with systemic lupus erythematosus and inactive disease
|Yates J,Whittington A,Mitchell P,Lechler RI,Lightstone L,Lombardi G||Clinical and experimental immunology (153:44)||2008|
|Not Applicable||1 µg/ml||
Insulin-like growth factor-I stimulates IL-10 production in human T cells.
M551 was used in ELISA to explore the immunomodulatory effects of insulin-like growth factor through regulation of cytokine production
|Kooijman R,Coppens A||Journal of leukocyte biology (76:862)||2004|