Mouse TIM-1/KIM-1/HAVCR Quantikine ELISA Kit

  (3 citations)     
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Assay Procedure
Citations (3)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (10 uL)
  • Sensitivity
    2.18 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant mouse TIM-1. The antibody components of this kit were raised against the short splice form of mouse TIM-1 (isoform 2) which has a 23 amino acid deletion following Pro182. This immunoassay detects both the short and long forms of mouse TIM-1.
  • 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 TIM-1 Immunoassay is a 3.5 hour solid phase ELISA designed to measure mouse TIM-1 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse TIM-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse TIM-1 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse TIM-1.

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, EDTA Plasma, Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 24.2 61.8 207 23.2 60 190
Standard Deviation 0.8 1.6 6.8 2.5 4.2 13.2
CV% 3.3 2.6 3.3 10.8 7 6.9

Recovery

The recovery of TIM-1 spiked into various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 100 91-106
EDTA Plasma (n=4) 89 88-91
Heparin Plasma (n=4) 87 86-89
Serum (n=4) 95 93-97
Urine (n=4) 102 94-115
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse TIM-1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 TIM-1/KIM-1/HAVCR [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TIM-1/KIM-1/HAVCR
TIM-1 (T cell immunoglobulin and mucin domain 1), also known as KIM-1 and HAVcr1, is expressed on many immune cell types and epithelial cells. It binds to phosphatidylserine (PS), LMIR5/CD300b, TIM-1 (homophilic), TIM-4, IgA, and the glycoproteins of a number of enveloped viruses. Its interaction with PS enables TIM-1 to mediate the phagocytosis and clearance of apoptotic cells. TIM-1 signaling costimulates T cell activation and enhances Th2 cytokine production. TIM-1 serves as a cellular entry receptor for hepatitis A virus, Ebolavirus and Marburgvirus. Polymorphisms are associated with susceptibility to atopy, autoimmunity, and severe hepatitis A virus infection in humans. A soluble form of TIM-1 is elevated in the urine during nephropathy.
  • Long Name:
    T Cell Immunoglobulin Mucin-1
  • Entrez Gene IDs:
    26762 (Human); 171283 (Mouse); 286934 (Rat); 102141332 (Cynomolgus Monkey)
  • Alternate Names:
    CD365; HAVCR1; HAVCR-1; HAVCRT cell immunoglobin domain and mucin domain protein 1; hepatitis A virus cellular receptor 1; Kidney injury molecule 1; KIM1; KIM-1; T-cell immunoglobulin and mucin domain-containing protein 1; TIM1; TIM-1; TIM-1TIM; TIM1TIMD-1; TIMD1T-cell membrane protein 1
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

The conjugate must remain at 2-8 °C during use. Bring all other 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 4 times for a total of 5 washes.

  8. 100 µL cold Conjugate
  9.   Add 100 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour on the benchtop.
  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 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:

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.

3 Citations: Showing 1 - 3
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Species
Sample Type
  1. Ameliorative Effect of Daidzein on Cisplatin-Induced Nephrotoxicity in Mice via Modulation of Inflammation, Oxidative Stress, and Cell Death
    Authors: H Meng, G Fu, J Shen, K Shen, Z Xu, Y Wang, B Jin, H Pan
    Oxid Med Cell Longev, 2017;2017(0):3140680.
    Species: Mouse
    Sample Type: Urine
  2. Human Alpha-1-Antitrypsin (hAAT) therapy reduces renal dysfunction and acute tubular necrosis in a murine model of bilateral kidney ischemia-reperfusion injury
    Authors: N Maicas, J van der Vl, J Bublitz, S Florquin, M Bakker-van, CA Dinarello, V Verweij, R Masereeuw, LA Joosten, LB Hilbrands
    PLoS ONE, 2017;12(2):e0168981.
    Species: Mouse
    Sample Type: Plasma
  3. Senescence marker protein-30/gluconolactonase deficiency exacerbates diabetic nephropathy through tubular injury in a mouse model of type 1 diabetes.
    Authors: Okada H, Senmaru T, Fukui M, Kondo Y, Ishigami A, Maruyama N, Obayashi H, Yamazaki M, Nakamura N, Hasegawa G
    J Diabetes Investig, 2015;6(1):35-43.
    Species: Mouse
    Sample Type: Urine

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