Mouse IL-34 ELISA Kit - Quantikine

Catalog # Availability Size / Price Qty
M3400
Mouse IL-34 ELISA Standard Curve
2 Images
Product Details
Procedure
Citations (4)
FAQs
Supplemental Products
Reviews (1)

Mouse IL-34 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
15.6 pg/mL
Assay Range
39.1 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse IL-34.
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 Mouse IL-34 immunoassay is a 4.5 hour solid-phase ELISA designed to measure IL-34 in mouse cell culture supernates, tissue lysates, serum, and plasma. It contains NS0-expressed recombinant mouse IL-34 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-34. Results obtained using natural mouse IL-34 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-34.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 99.6 273 870 107 284 903
Standard Deviation 2.4 4 12.7 8.5 15.5 33.3
CV% 2.4 1.5 1.5 7.9 5.5 3.7

Recovery

The recovery of IL-34 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 92-103
EDTA Plasma (n=4) 100 85-109
Heparin Plasma (n=4) 103 99-110
Serum (n=4) 100 95-107

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of IL-34 in each matrix were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse IL-34 ELISA Linearity

Scientific Data

Mouse IL-34 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.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-34

Interleukin 34 (IL-34; also known as uncharacterized protein C16orf77) is secreted as a homodimer consisting of 39 kDa monomers. It belongs to no known cytokine family. Human IL-34 is synthesized as a 242 amino acid (aa) precursor that contains a 20 aa signal sequence and a 222 aa mature chain. The mature chain contains one potential site of N-linked glycosylation. Human IL-34 is 71% identical to mouse IL-34 on the amino acid level.

IL-34 is expressed in various tissues, including the heart, brain, liver, kidney, spleen, thymus, testes, ovary, small intestine, prostate, and colon, and is most abundant in the spleen. The receptor for IL-34 is colony-stimulating factor 1 receptor (CSF-1R). IL-34 stimulates monocyte proliferation. In functional studies, IL-34, like CSF-1, the other ligand for CSF-1R, stimulated phosphorylation of extracellular signal-regulated kinase-1 and -2 (ERK1/2) in human monocytes. In addition, IL-34 promoted the formation of the colony-forming unit-macrophage (CFU-M), a macrophage progenitor, in human bone marrow cultures.

Long Name:
Interleukin 34
Entrez Gene IDs:
146433 (Human); 76527 (Mouse)
Alternate Names:
C16orf77; IL34; IL-34; interleukin 34
&#9888; 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.

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 on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 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 on the shaker.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. 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.

Citations for Mouse IL-34 Quantikine ELISA Kit

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.

4 Citations: Showing 1 - 4
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  1. CSF1R Ligands Expressed by Murine Gliomas Promote M-MDSCs to Suppress CD8+ T Cells in a NOS-Dependent Manner
    Authors: Takacs, GP;Garcia, JS;Hodges, CA;Kreiger, CJ;Sherman, A;Harrison, JK;
    Cancers
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Glioma-derived M-CSF and IL-34 license M-MDSCs to suppress CD8+ T cells in a NOS-dependent manner
    Authors: Takacs, GP;Garcia, JS;Hodges, CA;Kreiger, CJ;Sherman, A;Harrison, JK;
    bioRxiv : the preprint server for biology
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. Reno-protective effect of IL-34 inhibition on cisplatin-induced nephrotoxicity in mice
    Authors: Y Wada, M Iyoda, K Matsumoto, T Suzuki, S Tachibana, N Kanazawa, H Honda
    PLoS ONE, 2021-01-11;16(1):e0245340.
    Species: Mouse
    Sample Types: Serum
  4. M-CSF Mediates Host Defense during Bacterial Pneumonia by Promoting the Survival of Lung and Liver Mononuclear Phagocytes
    J Immunol, 2016-05-04;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates

FAQs

  1. Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?

    • A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

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Mouse IL-34 Quantikine ELISA Kit
By Anonymous on 03/06/2020
Sample Tested: Supernatant from mouse pancreatic cancer tumor spheres