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Human G-CSF DuoSet ELISA

Catalog #: DY214
21 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY214
DY214-05
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human G-CSF ELISA Standard Curve
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Product Details
Procedure
Citations (21)
FAQs
Supplemental Products
Reviews (1)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human G-CSF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

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 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

Kit Content

  • 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)

 

Scientific Data

N/A Human G-CSF ELISA Standard Curve View Larger

Human G-CSF ELISA Standard Curve

Product Datasheets

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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
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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human G-CSF DuoSet ELISA

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.

21 Citations: Showing 1 - 10
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  1. Ventilatory support and inflammatory peptides in hospitalised patients with COVID-19: A prospective cohort trial
    Authors: Gysan, MR;Milacek, C;Bal, C;Zech, A;Brugger, J;Milos, RI;Antoniewicz, L;Idzko, M;Gompelmann, D;
    PloS one
    Species: Human
    Sample Types: Serum
  2. Hypoxia Preconditioned Serum (HPS) Promotes Proliferation and Chondrogenic Phenotype of Chondrocytes In Vitro
    Authors: Jiang, J;Altammar, J;Cong, X;Ramsauer, L;Steinbacher, V;Dornseifer, U;Schilling, AF;Machens, HG;Moog, P;
    International journal of molecular sciences
    Species: Human
    Sample Types: Serum
  3. c-Met Mediated Cytokine Network Promotes Brain Metastasis of Breast Cancer by Remodeling Neutrophil Activities
    Authors: Liu, Y;Smith, MR;Wang, Y;D'Agostino, R;Ruiz, J;Lycan, T;Kucera, GL;Miller, LD;Li, W;Chan, MD;Farris, M;Su, J;Song, Q;Zhao, D;Chandrasekaran, A;Xing, F;
    Cancers
  4. DRP1-Mediated Mitochondrial Fission Regulates Lung Epithelial Response to Allergen
    Authors: SR Bruno, A Kumar, ZF Mark, R Chandrasek, E Nakada, N Chamberlai, B Mihavics, J Walzer, J Cahoon, AE Dixon, B Cunniff, V Anathy
    International Journal of Molecular Sciences, 2021-10-15;22(20):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Neutrophil function and bactericidal activity against Staphylococcus aureus after cardiac surgery with cardiopulmonary bypass
    Authors: M Lesouhaiti, M Gregoire, A Gacouin, V Coirier, A Frerou, C Piau, V Cattoir, E Dumontet, M Revest, P Tattevin, A Roisne, JP Verhoye, E Flecher, Y Le Tulzo, K Tarte, JM Tadié
    Journal of leukocyte biology, 2021-08-23;0(0):.
    Species: Human
    Sample Types: Plasma
  6. Characterization of low-density granulocytes in COVID-19
    Authors: LE Cabrera, PT Pekkarinen, M Alander, KHA Nowlan, NA Nguyen, S Jokiranta, S Kuivanen, A Patjas, S Mero, SH Pakkanen, S Heinonen, A Kantele, O Vapalahti, E Kekäläinen, T Strandin
    PloS Pathogens, 2021-07-06;17(7):e1009721.
    Species: Human
    Sample Types: Serum
  7. The dynamics of human bone marrow adipose tissue in response to feeding and fasting
    Authors: PK Fazeli, MA Bredella, OG Pachon-Peñ, W Zhao, X Zhang, AT Faje, M Resulaj, SP Polineni, TM Holmes, H Lee, EK O'Donnell, OA MacDougald, MC Horowitz, CJ Rosen, A Klibanski
    JCI Insight, 2021-06-22;0(0):.
    Species: Human
    Sample Types: Serum
  8. A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy
    Authors: J Li, A Wuethrich, AAI Sina, HH Cheng, Y Wang, A Behren, PN Mainwaring, M Trau
    Nature Communications, 2021-02-17;12(1):1087.
    Species: Human
    Sample Types: Serum
  9. Gene expression network analyses during infection with virulent and avirulent Trypanosoma cruzi�strains unveil a role for fibroblasts in neutrophil recruitment and activation
    Authors: AER Oliveira, MCA Pereira, AT Belew, LRP Ferreira, LMN Pereira, EGA Neves, MDCP Nunes, BA Burleigh, WO Dutra, NM El-Sayed, RT Gazzinelli, SMR Teixeira
    PLoS Pathog., 2020-08-18;16(8):e1008781.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Interleukin (IL)-17/IL-36 axis participates to the crosstalk between endothelial cells and keratinocytes during inflammatory skin responses
    Authors: L Mercurio, CM Failla, L Capriotti, C Scarponi, F Facchiano, M Morelli, S Rossi, G Pagnanelli, C Albanesi, A Cavani, S Madonna
    PLoS ONE, 2020-04-30;15(4):e0222969.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Probiotic Lactobacillus rhamnosus GR-1 is a unique prophylactic agent that suppresses infection-induced myometrial cell responses
    Authors: B Kim, O Shynlova, S Lye
    Sci Rep, 2019-03-18;9(1):4698.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Regulation of CXCL1 chemokine and CSF3 cytokine levels in myometrial cells by the MAFF transcription factor
    Authors: J Saliba, B Coutaud, V Solovieva, F Lu, V Blank
    J. Cell. Mol. Med., 2019-01-22;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Interleukin-1 primes human mesenchymal stem cells towards an anti-inflammatory and pro-trophic phenotype in vitro
    Authors: E Redondo-Ca, C Cunningham, J Miller, L Martuscell, S Aoulad-Ali, NJ Rothwell, CM Kielty, SM Allan, E Pinteaux
    Stem Cell Res Ther, 2017-04-17;8(1):79.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. IL-17 Promotes Neutrophil-Mediated Immunity by Activating Microvascular Pericytes and Not Endothelium
    J Immunol, 2016-08-17;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Opposite cytokine synthesis by fibroblasts in contact co-culture with osteosarcoma cells compared with transwell co-cultures.
    Authors: David M, Kelly E, Zoellner H
    Cytokine, 2013-03-19;62(1):48-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. IL-17C regulates the innate immune function of epithelial cells in an autocrine manner.
    Authors: Ramirez-Carrozzi V, Sambandam A, Luis E
    Nat. Immunol., 2011-10-12;12(12):1159-66.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Novel markers of inflammation identified in tumor necrosis factor receptor-associated periodic syndrome (TRAPS) by transcriptomic analysis of effects of TRAPS-associated tumor necrosis factor receptor type I mutations in an endothelial cell line.
    Authors: Rebelo SL, Amel-Kashipaz MR, Radford PM, Bainbridge SE, Fiets R, Fang J, McDermott EM, Powell RJ, Todd I, Tighe PJ
    Arthritis Rheum., 2009-01-01;60(1):269-80.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling.
    Authors: Weindl G, Naglik JR, Kaesler S, Biedermann T, Hube B, Korting HC, Schaller M
    J. Clin. Invest., 2007-12-01;117(12):3664-3672.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Role of mesothelial cell-derived granulocyte colony-stimulating factor in interleukin-17-induced neutrophil accumulation in the peritoneum.
    Authors: Witowski J, Ksiazek K, Warnecke C, Kuzlan M, Korybalska K, Tayama H, Wisniewska-Elnur J, Pawlaczyk K, Trominska J, Breborowicz A, Jorres A
    Kidney Int., 2007-01-17;71(6):514-25.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Prostanoid receptor expression by human airway smooth muscle cells and regulation of the secretion of granulocyte colony-stimulating factor.
    Authors: Clarke DL, Belvisi MG, Smith SJ, Hardaker E, Yacoub MH, Meja KK, Newton R, Slater DM, Giembycz MA
    Am. J. Physiol. Lung Cell Mol. Physiol., 2005-02-01;288(2):L238-50.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Toxoplasma gondii induces granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor secretion by human fibroblasts: implications for neutrophil apoptosis.
    Authors: Channon JY, Miselis KA, Minns LA, Dutta C, Kasper LH
    Infect. Immun., 2002-11-01;70(11):6048-57.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human G-CSF DuoSet ELISA
By Anonymous on 02/17/2020
Sample Tested: K562 human chronic myelogenous leukemia cell line