KIM1 ELISA Kit (Rat) (OKBB00354)

Catalog No: OKBB00354 (Formerly GWB-ZZD121 )

Size:96 Tests

Rat KIM-1 ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Rat
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's rat KIM1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for KIM1 has been precoated onto 96-well plates. Standards(NS0, S18-V238) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for KIM1 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the rat KIM1 amount of sample captured in plate.
• ELISA Kit Range: 31.2 pg/ml - 2,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	276	662	1186	359	710	1332
  St. Dev.	11.6	35.1	41.51	22.26	48.28	75.9
  %CV	4.20	5.30	3.5	6.20	6.8	5.69

• ELISA Kit Component:
  • Lyophilized recombinant mouse C-MET standard: 10 ng/tube x 2
  • Anti-mouse C-MET Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse C-MET antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 31.2pg/ml-2000pg/ml
  Principle: Aviva’s Rat KIM1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Rat KIM1 specific-specific polyclonal antibodies were precoated onto 96-well plates. The Rat specific detection monoclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Rat KIM1 amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before using.
  Background: KIM1（TIM-1）， also known as Hepatitis A virus cellular receptor 1, is a protein that in Rats is encoded by the HAVCR1 gene.[1][2][3] Infection of canine osteogenic sarcoma cells expressing HAVCR1 with HAV led Feigelstock et al. (1998) to conclude that the protein is indeed a receptor for the virus. Immunofluorescence microscopy demonstrated internalization of HAV by dog cells expressing HAVCR1. Using a monoclonal antibody to Rat Tim1, Umetsu et al. (2005) showed that Tim1 was expressed after activation of naive T cells and on T cells differentiated in Th2-polarizing conditions. By homology of synteny with the Rat Tim1 gene and database analysis, McIntire et al. (2001) mapped the HAVCR1 gene to 5q33.2. The standard used in this kit is recombinant Rat KIM-1, constituting from Y22-T212 amino acids, and 191amino acids with the molecular mass of 21.8KDa.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum, plasma (heparin, EDTA) and urine
• Sensitivity: <2 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Feigelstock D, Thompson P, Mattoo P, Zhang Y, Kaplan GG (Aug 1998). "The Rat homolog of HAVcr-1 codes
  for a hepatitis A virus cellular receptor". J Virol 72 (8): 6621–8.
  2. McIntire JJ, Umetsu SE, Akbari O, Potter M, Kuchroo VK, Barsh GS, Freeman GJ, Umetsu DT, DeKruyff RH
  (Nov 2001). "Identification of Tapr (an airway hyperreactivity regulatory locus) and the linked Tim gene family".
  Nat Immunol 2 (12): 1109–16. doi:10.1038/ni739..
  3. "Entrez Gene: HAVCR1 hepatitis A virus cellular receptor 1".
• Specificity: Natural and recombinant rat KIM1
• Immunogen: Expression system for standard: NS0; Immunogen sequence: S18-V238
• Description: For quantitative detection of rat KIM1 in cell culture supernatants, serum, plasma(heparin, EDTA) and urine.

Target Info

• Gene Symbol: HAVCR1
• Gene Full Name: hepatitis A virus cellular receptor 1
• Alias Symbols: HAVCR, HAVcr-1, Havcr1, HAVR1_HUMAN, Hepatitis A virus cellular receptor 1, Kidney injury molecule 1, KIM-1, T cell immunoglobin domain and mucin domain protein 1, T-cell immunoglobulin and mucin domain-containing protein 1, T-cell membrane protein 1, TIM, TIM-1, TIM1, TIMD-1, TIMD1
• NCBI Gene Id: 286934
• Protein Name: Hepatitis A virus cellular receptor 1
• Description of Target: KIM1(TIM-1), also known as Hepatitis A virus cellular receptor 1, is a protein that in Rat is encoded by the HAVCR1 gene. Infection of canine osteogenic sarcoma cells expressing HAVCR1 with HAV led to conclude that the protein is indeed a receptor for the virus. Immunofluorescence microscopy demonstrated internalization of HAV by dog cells expressing HAVCR1. Using a monoclonal antibody to rat Tim1, Tim1 was expressed after activation of naive T cells and on T cells differentiated in Th2-polarizing conditions. By homology of synteny with the rat Tim1 gene and database analysis, was mapped the HAVCR1 gene to 5q33.2.
• Uniprot ID: O54947
• Protein Accession #: NP_775172.1
• Nucleotide Accession #: NM_173149.2