Mouse IL-3 Quantikine ELISA Kit

R&D Systems | Catalog # M3000

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL)

Sensitivity

2.5 pg/mL

Assay Range

7.8-500 pg/mL (Cell Culture Supernates, Serum)
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Product Summary for Mouse IL-3 Quantikine ELISA Kit

The Quantikine Mouse IL-3 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-3 in cell culture supernates and mouse serum. It contains E. coli-expressed recombinant mouse IL-3 and antibodies raised against recombinant mouse IL-3. This Immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IL-3 showed dose 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 naturally occurring mouse IL-3.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Mouse

Specificity

Natural and recombinant mouse IL-3

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.

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) 33 119 396 32 120 400
Standard Deviation 1.7 8.3 22 2.5 5.5 18
CV% 5.2 7.0 5.6 7.8 4.6 4.5

Recovery for Mouse IL-3 Quantikine ELISA Kit

The recovery of mouse IL-3 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 104 91-114
Serum (n=5) 96 90-106

Linearity

To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IL-3 in each matrix were diluted with Calibrator Diluent and then assayed.

Mouse IL-3 ELISA Linearity

Scientific Data Images for Mouse IL-3 Quantikine ELISA Kit

Mouse IL-3 ELISA Standard Curve

Mouse IL-3 ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. 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: IL-3

Interleukin-3 (IL-3) is a secreted cytokine that stimulates the proliferation and differentiation of pluripotent hematopoietic stem cells and various lineage-committed progenitors. It also regulates the activation of mature mast cells, basophils, eosinophils, and macrophages. It is produced by activated T cells, thymic epithelial cells, activated mast cells, keratinocytes, neurons, and astrocytes. IL-3 signals through a receptor complex that contains IL-3 R alpha/CD123 and the common beta chain which is also a component of the receptors for IL-5 and GM-CSF. Receptors for IL-3 are present on bone marrow progenitors, macrophages, mast cells, eosinophils, megakaryocytes, basophils, and various myeloid leukemic cells. Human IL-3 does not show activity on mouse cells.

Long Name

Interleukin 3

Alternate Names

IL3, MCGF, MULTI-CSF

Entrez Gene IDs

3562 (Human); 16187 (Mouse); 24495 (Rat)

Gene Symbol

IL3

Additional IL-3 Products

Product Documents for Mouse IL-3 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 Mouse IL-3 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 Mouse IL-3 Quantikine ELISA Kit

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Protocols

View specific protocols for Mouse IL-3 Quantikine ELISA Kit (M3000):

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.

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