Rat GM-CSF Quantikine ELISA Kit

Discontinued Product

RGM00 has been discontinued.
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Rat GM-CSF Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL)
Sensitivity
2.1 pg/mL
Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum)
Specificity
Natural and recombinant rat GM-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 Rat GM-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat GM-CSF in cell culture supernates and rat serum. It contains E. coli-expressed recombinant rat GM-CSF and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant rat GM-CSF. Results obtained using natural rat GM-CSF showed dose response curves that were parallel to the standard curves obtained using the recombinant rat kit standards. These results indicate that this kit can be used to determine relative mass values for natural rat GM-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, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 55 176 746 56 172 723
Standard Deviation 4.6 6.9 30 5.5 10 43
CV% 8.4 3.9 4 9.8 5.8 5.9

Recovery

The recovery of rat GM-CSF spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=6) 97 89-107
Serum (n=14) 93 80-105

Linearity

To assess the linearity of the assay, six samples containing and/or spiked with high concentrations of rat GM-CSF in each matrix were diluted with Calibrator Diluent and then assayed.

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: GM-CSF

GM-CSF (Granulocyte-Macrophage Colony Stimulating Factor) induces the development of monocytes, neutrophils, eosinophils, and myeloid and dermal dendritic cells. It also acts as a neutrophil and dendritic cell chemoattractant. GM-CSF promotes Th1 and Th17 cell mediated autoimmune inflammation as well as the inflammatory activation of dendritic cells, microglia, alveolar macrophages, and eosinophils. In addition, it cooperates with G-CSF in promoting tumor cell proliferation and invasion. GM-CSF signals through a receptor complex composed of GM-CSF R alpha and the Common beta Chain (beta c) with Syndecan-2 as a potential co-receptor. The beta c subunit is shared by the receptor complexes for IL-3 and IL-5.

Long Name:
Granulocyte Macrophage Growth Factor
Entrez Gene IDs:
1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline)
Alternate Names:
colony stimulating factor 2 (granulocyte-macrophage); Colony-stimulating factor; CSF; CSF2; CSF-2; GMCSF; GM-CSF; GMCSFgranulocyte-macrophage colony-stimulating factor; granulocyte-macrophage colony stimulating factor; MGC131935; MGC138897; Molgramostim; molgramostin; Sargramostim

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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

FAQs

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

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