Human G-CSF Quantikine ELISA Kit

R&D Systems | Catalog # DCS50

R&D Systems
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Key Product Details

Assay Length

3.5 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Citrate Plasma (100 µL)

Sensitivity

20 pg/mL

Assay Range

39-2500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human G-CSF Quantikine ELISA Kit

The Quantikine Human G-CSF Immunoassay is a 3.5 or 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 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 kit standards. These results indicate that this kit can be used to determine relative mass values for natural human G-CSF.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

Sample Values

Serum/Plasma - Thirty-seven serum and plasma samples from apparently healthy volunteers were evaluated for the presence of human G-CSF in this assay. No medical histories were available for the donors used in this study. All samples measured less than the lowest human G-CSF standard, 39 pg/mL. 

Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate and stimulated with 10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and assayed for levels of natural human G-CSF.

ConditionDay 1 (pg/mL)Day 5 (pg/mL)
Unstimulated258118
Stimulated8541139

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 20 20 20
Mean (pg/mL) 177 515 1047 143 818 1514
Standard Deviation 4.5 9.4 22.0 17.0 72.0 77.1
CV% 2.5 1.8 2.1 11.9 8.8 5.1

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 280 827 1696 176 1094 2169
Standard Deviation 7.8 14.1 19.5 7.3 34.6 81.6
CV% 2.8 1.7 1.1 4.1 3.2 3.8

Recovery for Human G-CSF Quantikine ELISA Kit

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=5) 94 85-110
Citrate Plasma (n=5) 90 78-111
EDTA Plasma (n=5) 93 80-117
Heparin Plasma (n=5) 87 76-103
Serum (n=5) 88 75-106

Linearity

To assess the linearity of the assay, five 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 Images for Human G-CSF Quantikine ELISA Kit

Human G-CSF ELISA Cell Culture Supernate Standard Curve

Human G-CSF ELISA Cell Culture Supernate Standard Curve

Human G-CSF ELISA Serum/Plasma Standard Curve

Human G-CSF ELISA Serum/Plasma Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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

Alternate Names

C17orf33, CSF3, CSF3OS, Filgrastim, GCSF, Lenograstim, Pluripoietin

Entrez Gene IDs

1440 (Human); 12985 (Mouse)

Gene Symbol

CSF3

Additional G-CSF Products

Product Documents for Human G-CSF Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human G-CSF Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human G-CSF Quantikine ELISA Kit

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  • Human G-CSF Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: EDTA Plasma and Embryo tissue
    Verified Customer | Posted 02/06/2025
  • Human G-CSF Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 04/04/2018
    Human G-CSF Quantikine ELISA Kit DCS50
  • Human G-CSF Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: 4T1 mouse breast cancer cell line
    Verified Customer | Posted 04/03/2018
  • Human G-CSF Quantikine ELISA Kit
    Name: Jeff Pimentel
    Sample Tested: mouse serum
    Verified Customer | Posted 04/02/2018
  • Human G-CSF Quantikine ELISA Kit
    Name: Mike Ehrhardt
    Sample Tested: Serum
    Verified Customer | Posted 04/24/2017
    Human G-CSF Quantikine ELISA kit is used for Diagnostic use.
    Human G-CSF Quantikine ELISA Kit DCS50

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Protocols

View specific protocols for Human G-CSF Quantikine ELISA Kit (DCS50):

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.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  8.   Aspirate and wash 3 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    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.

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