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Human G-CSF Quantikine HS ELISA Kit

Catalog #: HSTCS0
8 Citations Datasheet / COA / SDS
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Control Products Available
QC97: Quantikine Immunoassay Control Set 1060 for HS Human G-CSF
Human G-CSF ELISA Calibrator Diluent RD5-5 Standard Curve
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Citations (8)
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human G-CSF Quantikine HS ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
Sensitivity
4.62 pg/mL
Assay Range
4.7 - 300 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human G-CSF
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine HS Human G-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure G-CSF in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human G-CSF and antibodies raised against the recombinant protein. It has been shown to accurately quantitate recombinant human G-CSF. Results obtained using natural human G-CSF showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that this kit can be used to determine relative mass values for natural human G-CSF.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 50.5 96.8 217 50.5 91.4 207
Standard Deviation 2.1 3.2 9.7 4 7 10.5
CV% 4.2 3.3 4.5 7.9 7.4 5.1

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 60 113 243 62.6 117 260
Standard Deviation 4.1 7.7 12.3 5.2 7.6 15.3
CV% 6.8 6.8 5.1 8.3 6.5 5.9

Recovery

The recovery of G-CSF spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 94-104
EDTA Plasma (n=4) 91 84-99
Heparin Plasma (n=4) 89 80-100
Serum (n=4) 88 80-94

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of G-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human G-CSF ELISA Linearity

Scientific Data

N/A Human G-CSF ELISA Calibrator Diluent RD5-5 Standard Curve View Larger

Human G-CSF ELISA Calibrator Diluent RD5-5 Standard Curve

N/A Human G-CSF ELISA Calibrator Diluent RD6P (diluted 1:2) Standard Curve View Larger

Human G-CSF ELISA Calibrator Diluent RD6P (diluted 1:2) Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: G-CSF

Granulocyte-colony stimulating factor (G-CSF) is a 24-25 kDa monomeric glycoprotein that regulates the proliferation, differentiation, and activation of hematopoietic cells in the neutrophilic granulocyte lineage. Mature human G-CSF is a 178 amino acid (aa) O-glycosylated protein that contains two intrachain disulfide bridges. In humans, alternate splicing generates a second minor isoform with a 3 aa deletion. Mouse and human G-CSF share 76% aa sequence identity, and the two proteins show species cross-reactivity. G-CSF is produced by activated monocytes and macrophages, fibroblasts, endothelial cells, astrocytes, neurons, and bone marrow stroma cells. In addition, various tumor cells express G-CSF constitutively.   


Human G-CSF receptor (G-CSF R) is a 120 kDa type I transmembrane glycoprotein that belongs to the hematopoietin receptor superfamily. The mature protein consists of a 603 aa extracellular domain (ECD), a 23 aa transmembrane segment, and a 186 aa cytoplasmic domain. The ECD contains an N-terminal Ig-like domain, a cytokine receptor homology domain, and three fibronectin type III domains. Alternate splicing of human G-CSF R generates additional isoforms including a potentially soluble form of the receptor. The ECDs of mouse and human G-CSF R share 63% aa sequence identity. G-CSF R forms a complex with the ligand in a 2:2 ratio. It is expressed on monocytes, neutrophils, megakaryocytes, platelets, myeloid progenitors, trophoblasts and placenta, endothelial cells, and various tumor cell types.   

G-CSF is an important regulator for granulopoiesis in vivo, and mutations in G-CSF R are associated with congenital neutropenia. G-CSF can support the growth of multilineage hematopoietic progenitor cells and mobilize them from the bone marrow into the bloodstream. G-CSF enhances the functional capacity of mature neutrophils and supports their survival by limiting the rate of apoptosis. G-CSF also enhances M-CSF induced monocytopoiesis from hematopoietic progenitor cells and stimulates the proliferation of peripheral Th2-inducing dendritic cells (30, 31). It promotes the development of T cell immune tolerance as well as tissue recovery following myocardial infarction and cerebral ischemia.

Long Name:
Granulocyte Colony Stimulating Factor
Entrez Gene IDs:
1440 (Human); 12985 (Mouse)
Alternate Names:
C17orf33; chromosome 17 open reading frame 33; colony stimulating factor 3 (granulocyte); CSF3; CSF3OS; Filgrastim; GCSF; G-CSF; GCSFlenograstim; granulocyte colony-stimulating factor; Lenograstim; MGC45931; Pluripoietin
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal microplate shaker.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. PROTECT FROM LIGHT.
  9.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 20 minutes.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human G-CSF Quantikine HS ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. Modulating bone marrow hematopoietic lineage potential to prevent bone metastasis in breast cancer
    Authors: JM Ubellacker, N Baryawno, N Severe, MJ DeCristo, J Sceneay, JN Hutchinson, MT Haider, CS Rhee, Y Qin, WM Gregory, AC Garrido-Ca, I Holen, JE Brown, RE Coleman, DT Scadden, SS McAllister
    Cancer Res., 2018-07-31;0(0):.
    Species: Human
    Sample Types: Plasma
  2. Differential expression of cell cycle and WNT pathway-related genes accounts for differences in the growth and differentiation potential of Wharton's jelly and bone marrow-derived mesenchymal stem cells
    Authors: AK Batsali, C Pontikoglo, D Koutroulak, KI Pavlaki, A Damianaki, I Mavroudi, K Alpantaki, E Kouvidi, G Kontakis, HA Papadaki
    Stem Cell Res Ther, 2017-04-26;8(1):102.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Phase I trial of combretastatin A4 phosphate (CA4P) in combination with bevacizumab in patients with advanced cancer.
    Clin. Cancer Res., 2012-05-29;18(12):3428-39.
    Species: Human
    Sample Types: Plasma
  4. Endogenous granulocyte colony-stimulating factor: a biomarker in acute ischemic stroke.
    Authors: Yu SC, Kuo CL, Huang CS, Chang CS, Wu SL, Su SL, Liu CS
    Biomarkers, 2012-03-23;17(4):319-24.
    Species: Human
    Sample Types: Plasma
  5. Rapid chemotherapy-induced acute endothelial progenitor cell mobilization: implications for antiangiogenic drugs as chemosensitizing agents.
    Authors: Shaked Y, Henke E, Roodhart JM, Mancuso P, Langenberg MH, Colleoni M, Daenen LG, Man S, Xu P, Emmenegger U, Tang T, Zhu Z, Witte L, Strieter RM, Bertolini F, Voest EE, Benezra R, Kerbel RS
    Cancer Cell, 2008-09-09;14(3):263-73.
    Species: Human
    Sample Types: Plasma
  6. Duffy (Fy), DARC, and neutropenia among women from the United States, Europe and the Caribbean.
    Authors: Grann VR, Ziv E, Joseph CK, Neugut AI, Wei Y, Jacobson JS, Horwitz MS, Bowman N, Beckmann K, Hershman DL
    Br. J. Haematol., 2008-08-15;143(2):288-93.
    Species: Human
    Sample Types: Plasma
  7. Neutropenia in 6 ethnic groups from the Caribbean and the U.S.
    Authors: Grann VR, Bowman N, Joseph C, Wei Y, Horwitz MS, Jacobson JS, Santella RP, Hershman DL
    Cancer, 2008-08-15;113(4):854-60.
    Species: Human
    Sample Types: Plasma
  8. Duffy antigen modifies the chemokine response in human endotoxemia.
    Authors: Mayr FB, Spiel AO, Leitner JM, Firbas C, Kliegel T, Jilma-Stohlawetz P, Derendorf H, Jilma B
    Crit. Care Med., 2008-01-01;36(1):159-65.
    Species: Human
    Sample Types: Plasma

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