Mouse G-CSF DuoSet ELISA

Catalog # Availability Size / Price Qty
DY414
DY414-05
Ancillary Products Available
Mouse G-CSF ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (21)
FAQs
Supplemental Products
Reviews

Mouse G-CSF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five and 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 mouse 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.

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)

Data Example

Mouse G-CSF ELISA Standard Curve

Product Datasheets

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

Mouse granulocyte-colony stimulating factor (G-CSF) is a 24-25 kDa monomeric glycoprotein that regulates the proliferation, differentiation and activation of hematopoietic cells. Mouse G-CSF cDNA encodes a 208 amino acid (aa) precursor protein with a 30 aa signal sequence that is proteolytically cleaved to form a 178 aa O-glycosylated mature protein containing two intrachain disulfide bridges. In humans, two distinct cDNA clones, encoding a 204 aa form and a minor alternatively spliced 207 aa form of G-CSF precursors, have been isolated. Mouse and human G-CSF are 76% identical at the aa sequence level and the two proteins show species cross-reactivity. G-CSF is produced primarily by monocytes and macrophages upon activation by endotoxin, TNF-alpha or IL-1. Other cell types, including fibroblasts, endothelial cells, astrocytes and bone marrow stroma cells, can also secrete G-CSF after activation. In addition, various tumor cells express G-CSF constitutively.   


Mouse G-CSF receptor (G-CSF R) is a 120 kDa, type I transmembrane glycoprotein that belongs to the hematopoietin receptor superfamily. The mature protein is 812 amino acids in length and contains a 601 aa extracellular region, a 24 aa transmembrane segment, and a 187 aa cytoplasmic domain. The extracellular region contains multiple modules, including an N-terminal Ig-like domain, a cytokine receptor homology domain, and three fibronectin type III domains. Based on crystallographic study, G-CSF receptor forms a complex with the ligand in a 2:2 ratio. Mouse and human G-CSF receptor share 63% aa sequence identity. Cells known to express G-CSF R include monocytes and neutrophils, megakaryocytes and platelets, CD34+ CD33+ and CD34+ CD38+ hematopoietic progenitors, trophoblasts, endothelial cells and various tumor cell types.    

G-CSF is an important regulator for granulopoiesis in vivo. It has been demonstrated that G-CSF can support the growth of multi-lineage hematopoietic progenitor cells without influencing their commitment to the myeloid lineage and mobilize hematopoietic progenitor cells from the bone marrow into the bloodstream. On mature neutrophils, G-CSF may regulate neutrophil survival by controlling their rate of apoptosis. G-CSF has also been shown to enhance the functional capacity of mature neutrophils. As a consequence of its effects on hematopoietic progenitor cells, G-CSF has been shown to enhance monocytopoiesis in the presence M-CSF. Within the peripheral blood stem cell population mobilized with G-CSF, selective increases in the number of T helper 2-inducing dendritic cells are found.

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, 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 Mouse 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
Filter your results:

Filter by:

  1. Keratinocyte-derived I&kappaB? drives psoriasis and associated systemic inflammation
    Authors: S Lorscheid, A Müller, J Löffler, C Resch, P Bucher, FC Kurschus, A Waisman, K Schäkel, S Hailfinger, K Schulze-Os, D Kramer
    JCI Insight, 2019;4(22):.
    Species: Mouse
    Sample Types: Serum
  2. PASylation of IL-1 Receptor antagonist (IL-1Ra) retains IL-1 blockade and extends its duration in mouse urate crystal-induced peritonitis
    Authors: NE Powers, B Swartzwelt, C Marchetti, DM de Graaf, A Lerchner, M Schlapschy, R Datar, U Binder, CK Edwards, A Skerra, CA Dinarello
    J. Biol. Chem., 2019;0(0):.
    Species: Mouse
    Sample Types: Plasma
  3. Breast and pancreatic cancer interrupt IRF8-dependent dendritic cell development to overcome immune surveillance
    Authors: MA Meyer, JM Baer, BL Knolhoff, TM Nywening, RZ Panni, X Su, KN Weilbaeche, WG Hawkins, C Ma, RC Fields, DC Linehan, GA Challen, R Faccio, RL Aft, DG DeNardo
    Nat Commun, 2018;9(1):1250.
    Species: Mouse
    Sample Types: Serum
  4. Modulation of Myelopoiesis Progenitors Is an Integral Component of Trained Immunity
    Authors: I Mitroulis, K Ruppova, B Wang, LS Chen, M Grzybek, T Grinenko, A Eugster, M Troullinak, A Palladini, I Kourtzelis, A Chatzigeor, A Schlitzer, M Beyer, LAB Joosten, B Isermann, M Lesche, A Petzold, K Simons, I Henry, A Dahl, JL Schultze, B Wielockx, N Zamboni, P Mirtschink, Ü Coskun, G Hajishenga, MG Netea, T Chavakis
    Cell, 2018;172(1):147-161.e12.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Secreted protein Del-1 regulates myelopoiesis in the hematopoietic stem cell niche
    Authors: I Mitroulis, LS Chen, RP Singh, I Kourtzelis, M Economopou, T Kajikawa, M Troullinak, A Ziogas, K Ruppova, K Hosur, T Maekawa, B Wang, P Subramania, T Tonn, P Verginis, M von Bonin, M Wobus, M Bornhäuser, T Grinenko, M Di Scala, A Hidalgo, B Wielockx, G Hajishenga, T Chavakis
    J. Clin. Invest., 2017;0(0):.
    Species: Mouse
    Sample Types: Plasma
  6. The Absence of NOD1 Enhances Killing of Aspergillus fumigatus Through Modulation of Dectin-1 Expression
    Authors: MS Gresnigt, M Jaeger, RK Subbarao M, O Rasid, G Jouvion, C Fitting, WJG Melchers, TD Kanneganti, A Carvalho, O Ibrahim-Gr, FL van de Vee
    Front Immunol, 2017;8(0):1777.
    Species: Mouse
    Sample Types: BALF
  7. Reducing hypoxia and inflammation during invasive pulmonary aspergillosis by targeting the Interleukin-1 receptor
    Sci Rep, 2016;6(0):26490.
    Species: Mouse
    Sample Types: Tissue Homogenates
  8. Epidermal RAF prevents allergic skin disease
    Elife, 2016;5(0):.
    Species: Mouse
    Sample Types: Serum
  9. The time-of-day of myocardial infarction onset affects healing through oscillations in cardiac neutrophil recruitment
    EMBO Mol Med, 2016;0(0):.
    Species: Mouse
    Sample Types: Plasma
  10. Extracellular Superoxide Dismutase Enhances the Recruitment of Immature Neutrophils to the Liver
    Infect Immun, 2016;0(0):.
    Species: Mouse
    Sample Types: Serum
  11. Infection-Mediated Priming of Phagocytes Protects against Lethal Secondary Aspergillus fumigatus Challenge
    Authors: A Savers, O Rasid, M Parlato, M Brock, G Jouvion, B Ryffel, JM Cavaillon, G Eberl, O Ibrahim-Gr
    PLoS ONE, 2016;11(4):e0153829.
    Species: Mouse
    Sample Types: Tissue Homogenates
  12. Thrombospondin-1 restrains neutrophil granule serine protease function and regulates the innate immune response during Klebsiella pneumoniae infection.
    Authors: Zhao Y, Olonisakin T, Xiong Z, Hulver M, Sayeed S, Yu M, Gregory A, Kochman E, Chen B, Mallampalli R, Sun M, Silverstein R, Stolz D, Shapiro S, Ray A, Ray P, Lee J
    Mucosal Immunol, 2015;8(4):896-905.
    Species: Mouse
    Sample Types: Tissue Homogenates
  13. BMP4 inhibits breast cancer metastasis by blocking myeloid-derived suppressor cell activity.
    Authors: Cao Y, Slaney C, Bidwell B, Parker B, Johnstone C, Rautela J, Eckhardt B, Anderson R
    Cancer Res, 2014;74(18):5091-102.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  14. A new protective role for S100A9 in regulation of neutrophil recruitment during invasive pneumococcal pneumonia.
    Authors: De Filippo K, Neill D, Mathies M, Bangert M, McNeill E, Kadioglu A, Hogg N
    FASEB J, 2014;28(8):3600-8.
    Species: Mouse
    Sample Types: Tissue Homogenates
  15. Regulation of adult hematopoiesis by the a disintegrin and metalloproteinase 10 (ADAM10).
    Authors: Weber S, Wetzel S, Prox J, Lehmann T, Schneppenheim J, Donners M, Saftig P
    Biochem Biophys Res Commun, 2013;442(3):234-41.
    Species: Mouse
    Sample Types: Plasma
  16. The interaction between C5a and both C5aR and C5L2 receptors is required for production of G-CSF during acute inflammation.
    Authors: Bosmann M, Haggadone M, Zetoune F, Sarma J, Ward P
    Eur J Immunol, 2013;43(7):1907-13.
    Species: Mouse
    Sample Types: Plasma
  17. Defective regulation of CXCR2 facilitates neutrophil release from bone marrow causing spontaneous inflammation in severely NF-kappa B-deficient mice.
    Authors: von Vietinghoff S, Asagiri M, Azar D
    J. Immunol., 2010;185(1):670-8.
    Species: Mouse
    Sample Types: Plasma
  18. IL-17A controls IL-17F production and maintains blood neutrophil counts in mice.
    Authors: von Vietinghoff S, Ley K
    J. Immunol., 2009;183(2):865-73.
    Species: Mouse
    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;71(6):514-25.
    Species: Mouse
    Sample Types: Peritoneal Lavage Fluid
  20. IL-23 is required for neutrophil homeostasis in normal and neutrophilic mice.
    Authors: Smith E, Zarbock A, Stark MA, Burcin TL, Bruce AC, Foley P, Ley K
    J. Immunol., 2007;179(12):8274-9.
    Species: Mouse
    Sample Types: Serum
  21. Choosing the frequency of deep inflation in mice: balancing recruitment against ventilator-induced lung injury.
    Authors: Allen GB, Suratt BT, Rinaldi L, Petty JM, Bates JH
    Am. J. Physiol. Lung Cell Mol. Physiol., 2006;291(4):L710-7.
    Species: Mouse
    Sample Types: BALF

FAQs

No product specific FAQs exist for this product, however you may

View all ELISA FAQs

Reviews for Mouse G-CSF DuoSet ELISA

There are currently no reviews for this product. Be the first to review Mouse G-CSF DuoSet ELISA and earn rewards!

Have you used Mouse G-CSF DuoSet ELISA?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review