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Human G-CSF DuoSet ELISA

R&D Systems, part of Bio-Techne | Catalog # DY214

R&D Systems, part of Bio-Techne
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DY214
DY214-05

Ancillary Products Available

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

31.2-2000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human G-CSF DuoSet ELISA Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Product Summary for Human G-CSF DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human G-CSF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Label

HRP

Scientific Data Images for Human G-CSF DuoSet ELISA

Human G-CSF ELISA Standard Curve

Human G-CSF ELISA Standard Curve

Kit Contents for Human G-CSF DuoSet ELISA

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

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 DuoSet ELISA

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human G-CSF DuoSet ELISA

For research use only

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