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Human TIM-1 / KIM-1 / HAVCR ELISA Standard Curve
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Human TIM-1/KIM-1/HAVCR DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sufficient Materials
For fifteen 96-well plates*
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 human T cell Immunoglobulin Mucin-1 (TIM-1). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, and urine samples. Diluents for complex matrices, such as serum, plasma, and urine, 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 Examples

Human TIM-1 / KIM-1 / HAVCR ELISA Standard Curve

Human TIM-1/KIM-1/HAVCR ELISA Standard Curve

Product Datasheets

Preparation and Storage

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: TIM-1/KIM-1/HAVCR

T cell immunoglobulin and mucin domain 1 (TIM-1), also known as Kidney injury molecule 1 (KIM-1) and Hepatitis A virus cellular receptor 1 (HAVcr1), is a member of the TIM family which is involved in the regulation of innate and adaptive immune responses (1, 2). TIM-1 is a type I transmembrane protein that contains an N-terminal immunoglobulin-like domain, a mucin domain with O- and N-linked glycosylations, a transmembrane segment, and a cytoplasmic signaling domain (3, 4). Multiple TIM-1 variants can be produced due to polymorphisms or alternative splicing resulting in deletions in the mucin domain (3). Some of these polymorphisms are associated with susceptibility to atopy, autoimmunity, and severe hepatitis A virus infection in humans (5). Within the extracellular domain, human TIM-1 shares 41% amino acid sequence identity with mouse and rat TIM-1. 

In vivo, TIM-1 is expressed on splenic B cells and is a marker for the identification of IL-10+ regulatory B cells (6, 7). TIM-1 is also expressed on CD4+ T cells, mast cells, invariant NKT (iNKT) cells, dendritic cells, kidney epithelium and a broad range of mucosal epithelium (4, 8-15). The expression of TIM-1 is upregulated on activated Th2 cells, after dendritic cell maturation, and on kidney tubular epithelial cells after injury (4, 9, 13, 14, 16, 17). Metalloproteinase-mediated cleavage of TIM-1 at the membrane-proximal region results in the release of a soluble form of TIM-1 which is detectable in the urine and in circulation (18, 19). Urinary TIM-1 is highly elevated in nephropathy and may be a useful biomarker for renal damage (16, 20 - 25). 
TIM-1 has been reported to be a receptor for a number of ligands, including phosphatidylserine, leukocyte mono-immunoglobulin-like receptor 5 (LMIR5/CD300b), TIM-1 (homophilic), TIM-4, IgA, and the glycoproteins of a number of enveloped viruses (5, 15, 26-33). Its interaction with phosphatidylserine enables TIM-1 to mediate the phagocytosis of apoptotic cells (26-28). In TIM-1-bearing iNKT cells, interaction with apoptotic cells can also result in iNKT cell activation, proliferation, and cytokine production (11). Interactions between cell-surface or soluble TIM-1 with LMIR5 is proposed to induce LMIR5-mediated activation of myeloid cells including macrophages/monocytes, mast cells, neutrophils, and dendritic cells (29). These interactions contribute to tissue homeostasis and damage during kidney injury (29). Ligandinduced TIM-1 signaling costimulates T cell activation and enhances Th2 cytokine production (9, 31, 34). In humans, TIM-1 serves as a cellular entry receptor for various viruses, including hepatitis A virus, Ebolavirus and Marburgvirus (15, 33).

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

Assay Procedure


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 Human TIM-1/KIM-1/HAVCR 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.

10 Citations: Showing 1 - 10
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  1. Early identification of acute kidney injury in Russell's viper (Daboia russelii) envenoming using renal biomarkers
    Authors: I Ratnayake, F Mohamed, NA Buckley, IB Gawaramman, DM Dissanayak, U Chathurang, M Munasinghe, K Maduwage, S Jayamanne, ZH Endre, GK Isbister
    PLoS Negl Trop Dis, 2019;13(7):e0007486.
    Species: Human
    Sample Types: Urine
  2. Acute Kidney Injury Biomarkers Predict an Increase in Serum Milrinone Concentration Earlier Than Serum Creatinine-Defined Acute Kidney Injury in Infants After Cardiac Surgery
    Authors: KM Gist, DS Cooper, J Wrona, S Faubel, C Altmann, Z Gao, BS Marino, J Alten, KM Hock, T Mizuno, AA Vinks, MS Joy, MF Wempe, MR Bennett, SL Goldstein
    Ther Drug Monit, 2018;40(2):186-194.
    Species: Human
    Sample Types: Urine
  3. Propofol-based anaesthesia versus sevoflurane-based anaesthesia for living donor kidney transplantation: results of the VAPOR-1 randomized controlled trial
    Authors: GJ Nieuwenhui, VB Nieuwenhui, MAJ Seelen, SP Berger, MC van den He, JGM Burgerhof, PJ Ottens, RJ Ploeg, HGD Leuvenink, MMRF Struys
    Br J Anaesth, 2017;118(5):720-732.
    Species: Human
    Sample Types: Urine
  4. Dexamethasone Modifies Cystatin C-Based Diagnosis of Acute Kidney Injury During Cisplatin-Based Chemotherapy
    Authors: TJ Pianta, JW Pickering, L Succar, M Chin, T Davidson, NA Buckley, F Mohamed, ZH Endre
    Kidney Blood Press. Res, 2017;42(1):62-75.
    Species: Human
    Sample Types: Urine
  5. Assessment of Worldwide Acute Kidney Injury, Renal Angina and Epidemiology in critically ill children (AWARE): study protocol for a prospective observational study.
    Authors: Basu R, Kaddourah A, Terrell T, Mottes T, Arnold P, Jacobs J, Andringa J, Goldstein S
    BMC Nephrol, 2016;16(0):24.
    Species: Human
    Sample Types: Urine
  6. Expression of Tim-1 in primary CNS lymphoma
    Cancer Med, 2016;5(11):3235-3245.
    Species: Human
    Sample Types: CSF
  7. Urinary adiponectin is an independent predictor of progression to end-stage renal disease in patients with type 1 diabetes and diabetic nephropathy.
    Authors: Panduru N, Saraheimo M, Forsblom C, Thorn L, Gordin D, Waden J, Tolonen N, Bierhaus A, Humpert P, Groop P
    Diabetes Care, 2015;38(5):883-90.
    Species: Human
    Sample Types: Urine
  8. Soluble TNF receptors and kidney dysfunction in the elderly.
    Authors: Carlsson A, Larsson T, Helmersson-Karlqvist J, Larsson A, Lind L, Arnlov J
    J Am Soc Nephrol, 2014;25(6):1313-20.
    Species: Human
    Sample Types: Urine
  9. Urinary L-FABP and anaemia: distinct roles of urinary markers in type 2 diabetes.
    Authors: von Eynatten M, Baumann M, Heemann U
    Eur. J. Clin. Invest., 2010;40(2):95-102.
    Species: Human
    Sample Types: Urine
  10. Assay validation for KIM-1: human urinary renal dysfunction biomarker.
    Authors: Chaturvedi S, Farmer T, Kapke GF
    Int. J. Biol. Sci., 2009;5(2):128-34.
    Species: Human
    Sample Types: Urine


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By Anonymous on 12/25/2018
Application: Sample Tested: Serum and Plasma

The only reason I didn't give the kit 5 stars is because we had a few technical issues with it that took some troubleshooting time to work out, but RnD Systems was very prompt in helping us resolve it and even sent a new kit after we found the problem.