Sandwich ELISA analysis of human GM-CSF was performed using a Human GM-CSF Colorimetric ELISA kit (Product # EHGMCSF) by loading 50 ul per well of a mouse anti-human GM-CSF biotinylated antibody (Product # M501B) at a concentration of 0.05 ug/ml and 50 ul of human GM-CSF Recombinant Protein (Product # SGMCSF) at 600, 240, 96, 38.4, 15.4, and 0 pg/ml (both in quadruplicate) across a 9 ug/ml mouse anti-human GM-CSF (Product # M500A) pre-coated plate. The plate was incubated for 3 hours at room temperature, washed, and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at 1:2000 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
|Tested species reactivity||Human|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Glycosylated, CHO-derived recombinant human GM-CSF|
|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.
M500A targets Human GM-CSF in ELISA, and WB applications and shows reactivity with Human samples.
The M500A immunogen is glycosylated, CHO-derived recombinant human GM-CSF.
M500A detects Human GM-CSF which has a predicted molecular weight of approximately 14 kDa.
The M500A GMCSF antibody (clone 3092) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M501B (biotinylated conjugate of clone 1089). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody M500A (clone 3092) and biotinylated antibody M501B (clone 1089) have successfully been used in combination with recombinant GMCSF protein SGMCSF in ELISA applications.
The protein encoded by this gene is a cytokine that controls the production, differentiation, and function of granulocytes and macrophages. The active form of the protein is found extracellularly as a homodimer. This gene has been localized to a cluster of related genes at chromosome region 5q31, which is known to be associated with interstitial deletions in the 5q- syndrome and acute myelogenous leukemia. Other genes in the cluster include those encoding interleukins 4, 5, and 13.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Borrelia burgdorferi infection regulates CD1 expression in human cells and tissues via IL1-ß.
M500A was used in ELISA to study the effect of Borrelia burgdorferi infection on CD1 expression
|Yakimchuk K,Roura-Mir C,Magalhaes KG,de Jong A,Kasmar AG,Granter SR,Budd R,Steere A,Pena-Cruz V,Kirschning C,Cheng TY,Moody DB||European journal of immunology (41:694)||2011|
|Not Applicable||Not Cited||
Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.
M500A was used in ELISA to test if factors released from human alveolar macrophages exposed to air pollution particles accelerate the maturation of granulocyte precursors
|Suwa T,Hogg JC,Vincent R,Mukae H,Fujii T,van Eeden SF||Experimental lung research (28:1)||2002|
Signal transduction of the human granulocyte-macrophage colony-stimulating factor and interleukin-3 receptors involves tyrosine phosphorylation of a common set of cytoplasmic proteins.
M500A was used in western blot to study the role of human GM-CSF and interleukin 3 receptor in phosphorylation of cytoplasmic proteins
|Kanakura Y,Druker B,Cannistra SA,Furukawa Y,Torimoto Y,Griffin JD||Blood (76:706)||1990|