Human Granulocyte-Macrophage Colony Stimulating Factor,GM-CSF ELISA Kit

Instructions
Code CSB-E04568h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Human Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) ELISA Kit is used for the quantitative measurement of human GM-CSF in many biological fluids, including serum, plasma, cell culture supernates, and tissue homogenates. GM-CSF, also known as CSF2, is a monomeric glycoprotein secreted by macrophages, T cells, mast cells, natural killer cells, endothelial cells, and fibroblasts. It is an important cytokine that exerts effects in both innate and adaptive immunity. Acting as an inducer, CSF2 stimulates myeloid progenitors to proliferate and differentiate into granulocyte and macrophage. CSF2 binding to its receptor CSF2R triggers four main signaling pathways, including JAK/STAT, PI3K-AKT, ERK1/2, and NF-kB pathways, thus promoting cell differentiation, survival, and inflammation.

The assay is based on the Sandwich ELISA mechanism and enzyme-substrate chromogenic reaction. The intensity of the color is proportional to the GM-CSF content in the sample. This kit has been verified with pg-level of sensitivity, high specificity, precision less than 10%, good linearity, high recovery, and lot-to-lot consistency. More validation details are described in the product instructions.

Target Name colony stimulating factor 2 (granulocyte-macrophage)
Alternative Names Colony stimulating factor 2 (granulocyte-macrophage) ELISA Kit; Colony Stimulating Factor 2 ELISA Kit; Colony stimulating factor ELISA Kit; Colony-stimulating factor ELISA Kit; CSF 2 ELISA Kit; CSF ELISA Kit; CSF2 ELISA Kit; CSF2_HUMAN ELISA Kit; GM-CSF ELISA Kit; GMCSF ELISA Kit; Granulocyte Macrophage Colony Stimulating Factor ELISA Kit; Granulocyte-macrophage colony-stimulating factor ELISA Kit; MGC131935 ELISA Kit; MGC138897 ELISA Kit; MGI1GM ELISA Kit; Molgramostin ELISA Kit; Pluripoietin-a ELISA Kit; Sargramostim ELISA Kit
Abbreviation CSF2
Uniprot No. P04141
Species Homo sapiens (Human)
Sample Types serum, plasma, cell culture supernates, tissue homogenates
Detection Range 15.6 pg/mL-1000 pg/mL
Sensitivity 3.9 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human GM-CSF in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)
1:1 Average % 89
Range % 82-99
1:2 Average % 95
Range % 88-102
1:4 Average % 95
Range % 86-99
1:8 Average % 95
Range % 90-105
Recovery
The recovery of human GM-CSF spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range
Serum (n=5) 92 86-100
EDTA plasma (n=4) 96 90-110
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/ml OD1 OD2 Average Corrected
1000 2.987 2.913 2.950 2.792
500 2.417 2.429 2.423 2.265
250 1.674 1.686 1.680 1.522
125 0.954 0.963 0.959 0.801
62.5 0.576 0.562 0.569 0.411
31.2 0.386 0.394 0.390 0.232
15.6 0.261 0.268 0.265 0.107
0 0.156 0.159 0.158  
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human GM-CSF. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled GM-CSF antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) ---Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Data

Function Cytokine that stimulates the growth and differentiation of hematopoietic precursor cells from various lineages, including granulocytes, macrophages, eosinophils and erythrocytes.
Gene References into Functions
  1. GM-CSF could be induced by M. pneumoniae infection in vivo and vitro. Childen with high level GM-CSF had longer duration of fever. GM-CSF probably plays a vital role in neutrophil inflammation in M. pneumoniae infection. PMID: 30007865
  2. Data show that CD4 T cells from synovial fluid are enriched for granulocyte-macrophage colony-stimulating factor (GM-CSF) production, suggests that the GM-CSF programme may be a primary pathogenic process. PMID: 29142230
  3. GM-CSF bioavailability plays a critical role in maintaining intestinal homeostasis. Decreased bioavailability coupled with the genetic risk markers and/or smoking results in aggressive ileal Crohn's disease behavior. PMID: 29434451
  4. Data show that GIF (ORF117 ) serves as a competitive decoy receptor by leveraging binding hotspots underlying the cognate receptor interactions of granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-2 (IL-2). PMID: 27819269
  5. As translated to human disease, these findings suggest vaccine-mediated expansion of GM-CSF-producing T cells could be an effective prophylactic or therapeutic TB strategy. PMID: 29233902
  6. In a subset of patients who received GM-CSF, circulating myeloid-derived suppressor cells (MDSC), and anti-GM-CSF-neutralizing antibodies (Nabs) were also modulated. The majority of patients developed anti-GM-CSF Nabs, which correlated with improved RFS and OS PMID: 28536308
  7. this study shows that cortisol inhibits CSF2 via DNA methylation and inhibits invasion in first-trimester trophoblast cells PMID: 28846166
  8. Oral and periodontal innate immunity is affected by HIV viremia and ART. GCF IL-8, G-CSF, as well as serum IL-8, MCP-1 and GM-CSF may be useful biomarkers for the detection of disease presence and/or its severity due to HIV infection and ART use. PMID: 28043030
  9. This review summarizes a number of findings to provide the currently available information regarding the anticancer immune response of GM-CSG. [review] PMID: 27364892
  10. Data suggest that the intratumoral GM-CSF expression, as a potentially independent prognostic biomarker for recurrence, might improve conventional clinical and pathologic analysis to refine outcome prediction for clinically localized clear-cell renal cell carcinoma (ccRCC) patients after surgery. PMID: 27016418
  11. High GM-CSF expression is associated with breast Cancer. PMID: 28754674
  12. In gastric cancer (GC), tumour-derived GM-CSF activated neutrophils and induced neutrophil PD-L1 expression via Janus kinase (JAK)-signal transducer and activator of transcription 3 (STAT3) signalling pathway. The activated PD-L1(+) neutrophils effectively suppressed normal T-cell immunity in vitro and contributed to the growth and progression of human GC in vivo. PMID: 28274999
  13. The IL-3/ GM-CSF effected on the myofibroblastic differentiation of human adipose derive stromal cells (hASCs) as well as it did on human dermal fibroblasts (HDFs). PMID: 28377320
  14. Obesity alters the lung neutrophil infiltration to enhance breast cancer metastasis through IL5 and GM-CSF. PMID: 28737771
  15. Data suggest that the methylotrophic yeast Pichia pastoris is an effective recombinant host for heterologous granulocyte-macrophage colony-stimulating factor (rhGM-CSF) production. PMID: 28376863
  16. Letter: Knockdown of either filaggrin or loricrin increases the productions of interleukin (IL)-1alpha, IL-8, IL-18 and granulocyte macrophage colony-stimulating factor in stratified human keratinocytes. PMID: 26381575
  17. Impaired RASGRF1/ERK-mediated GM-CSF response characterizes CARD9 deficiency in French-Canadians. PMID: 26521038
  18. Honokiol could possess potential anti-inflammatory effects and inhibits TNF-alpha-induced IL-1beta, GM-CSF and IL-8 production in PBMCs from rheumatoid arthritis patients. PMID: 26339358
  19. The canonical NFkappaB signaling in fibroblasts, but not in tumor cells, was shown to be responsible for the induced and constitutive CSF2 expression. PMID: 25919140
  20. GM-CSF primes IL-13 production by macrophages via PAR-2. PMID: 25633855
  21. analysis of an IL2RA polymorphism that controls GM-CSF production in human TH cells PMID: 25278028
  22. NFKBIZ gene knockdown in bronchial epithelial cells suppresses the release of IL-1b-induced IL-6 and GMCSF. PMID: 25629767
  23. CFH Y402H polymorphism is associated with elevated vitreal GM-CSF and choroidal macrophages in the postmortem human eye. PMID: 25814824
  24. Influenza infection stimulates the secretion of IL-8 and GM-CSF by alveolar epithelial cells. PMID: 26033355
  25. Periodic fever, aphthous stomatitis, pharyngitis and cervical adenopathy syndrome febrile episodes are characterized by activation of GM-CSF and IL-8 with Th1 skewing. PMID: 24670131
  26. activation of STAT3 induces the expression of GM-CSFRa that protects CLL cells from apoptosis, suggesting that inhibition of STAT3 or GM-CSFRa may benefit patients with chronic lymphocytic leukemia PMID: 24836891
  27. G-CSF- or GM-CSF-secreting cancers, albeit not very common are, however, among the most rapidly advancing ones due to a cytokine-mediated immune suppression and angiogenesis. [Review] PMID: 24692240
  28. Treatment with GM-CSF increased the expression of vimentin and ZEB1, similar to TGF-b1 treatment, and decreased the expression of E-cadherin. Our results suggest that GM-CSF could induce EMT in human HERS/ERM cells. PMID: 24258001
  29. GM-CSF polymorphism may be involved in the pathogenesis of atopic dermatitis. PMID: 24117406
  30. The association of GM-CSF expression with systemic inflammation highlights the potential role of Th17-related cytokines in the pathology of juvenile idiopathic arthritis. PMID: 24692225
  31. M-CSF induces macrophage transformation by upregulating c-Jun with a certain synergy of NFkappaB. PMID: 24100343
  32. The detection of basal level of IgM rheumatoid factor, IgM and also certain cytokines can be useful in prognosis of effectiveness of rituximab therapy under rheumatoid arthritis. PMID: 25080789
  33. Cytokines and signaling molecules predict clinical outcomes in sepsis. PMID: 24244449
  34. Granulocyte colony-stimulating factor produces long-term changes in gene and microRNA expression profiles in CD34+ cells from healthy donors. PMID: 24056818
  35. mesenchymal-like breast cancer cells activate macrophages to a tumor-associated macrophage-like phenotype by GM-CSF. PMID: 24823638
  36. The comparison between the cytokine profiles of ET and PV patients showed a statistically significant increase of IL-4, IL-8, GM-CSF, interferon -gamma, MCP-1, PDGF-BB, and VEGF in the ET group PMID: 24463275
  37. Data indicate that differentially expressed IL-17, IL-22, and IL-23 levels are associated with K-ras in a stage-specific fashion along colorectal cancer progression and an association was established between mutant K-ras and GM-CSF and IFN-gamma. PMID: 24040001
  38. in the type 2 diabetes mellitus group, significantly higher IL-4 level on the first and the third post dengue virus infection days, while higher IL-10 and granulocyte-macrophage colony-stimulating factor were detected on the third postinfection day. PMID: 24078930
  39. loss of BMPR2 induced prolonged phospho-p38 MAPK in response to TNF, and this increased GADD34-PP1 phosphatase activity, dephosphorylating eukaryotic translation initiation factor (eIF2alpha), and derepressing GM-CSF mRNA translation PMID: 24446489
  40. Since GM-CSF plays an important role during pregnancy, the reduced levels of GM-CSF in the cervico-vaginal fluid of human papillomavirus(+) women might contribute to explain the reduced live birth rate. PMID: 23920355
  41. SECTM1 secreted from bone marrow stromal cells may interact with CD7 to influence GM-CSF expression in leukemic cells. PMID: 24211252
  42. Describe GM-CSF ELISA for routine clinical diagnosis of autoimmune pulmonary alveolar proteinosis. PMID: 24275678
  43. The data presented suggest that -677C/A in the promoter region of the GM-CSF gene does not play a role in acne pathogenesis. PMID: 23069312
  44. CBL linker and RING finger mutants lead to enhanced GM-CSF signaling due to elevated kinase expression, which can be blocked using small molecule inhibitors targeting specific downstream pathways. PMID: 23696637
  45. impairment of NFATC2-calcineurin interaction promotes megakaryocyte proliferation through up-regulation of CSF2 transcription PMID: 22911897
  46. Overexpression of a nonphosphorylatable MT1-MMP mutant (Y573F) abrogates CSF-2 and CSF-3 transcriptional increases. PMID: 23548906
  47. GM-CSF contributes to aortic aneurysms resulting from SMAD3 deficiency PMID: 23585475
  48. Within pediatric Crohn disease, neutrophil GM-CSF-dependent STAT5 activation and bacterial killing were reduced as GM-CSF autoantibodies increased. PMID: 23600834
  49. This study demonistrated that GMCSF is downregulation in brain in patient with Alzheimer's disease. PMID: 22430742
  50. IL-27 suppresss GM-CSF production under nonpolarizing and T helper (Th)1 cell, but not T helper (Th)17 cell conditions, in both murine and human cultures. PMID: 22837488

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Subcellular Location Secreted
Protein Families GM-CSF family
Database Links

HGNC: 2434

OMIM: 138960

KEGG: hsa:1437

STRING: 9606.ENSP00000296871

UniGene: Hs.1349

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