Human Signal Transduction ELISA Kits
Human IGF2R (Insulin Like Growth Factor 2 Receptor) CLIA Kit (HUES00317)
- SKU:
- HUES00317
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 75pg/mL
- Range:
- 125-8000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- CD222, CIMPR, M6P-R, MPR1, MPRI, cation-independent mannose-6-phosphate receptor
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Signal Transduction
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 125.00-8000 pg/mL |
| Sensitivity: | 75.00 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Tissue homogenates,cell lysates and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human IGF2R in samples. No significant cross-reactivity or interference between Human IGF2R and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human IGF2R. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IGF2R and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IGF2R, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human IGF2R. The concentration of Human IGF2R in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | IGF2R: a multifunctional type I transmembrane protein receptor that binds insulin-like growth factor 2 at the cell surface and mannose-6-phosphate-tagged proteins in the trans-Golgi network. Also known as the cation-independent mannose-6 phosphate (M6P) receptor. Clears IGF2 from the cell surface to attenuate signaling, and transports M6P-tagged lysosomal enzymes from the Golgi complex and the cell surface to lysosomes. M6P-tagged lysosomal enzymes bind specifically to mannose-6- phosphate receptors in the Golgi apparatus and the resulting receptor-ligand complex is transported to an acidic prelyosomal compartment where the low pH mediates the dissociation of the complex. Acts as a positive regulator of T-cell coactivation, by binding DPP4. Belongs to the MRL1/IGF2R family. |
| UniProt Protein Details: | Protein type:Membrane protein, integral; Receptor, misc. ; Vesicle Chromosomal Location of Human Ortholog: 6q26 Cellular Component: cell surface; clathrin coat; early endosome; endocytic vesicle; endosome; extracellular space; focal adhesion; integral to plasma membrane; late endosome; lysosomal membrane; membrane; nuclear envelope lumen; perinuclear region of cytoplasm; trans-Golgi network; trans-Golgi network transport vesicle Molecular Function:enzyme binding; G-protein alpha-subunit binding; G-protein coupled receptor activity; glycoprotein binding; identical protein binding; insulin-like growth factor II binding; insulin-like growth factor receptor activity; mannose binding; phosphoprotein binding; protein binding; receptor activity; retinoic acid binding; transporter activity Biological Process: G-protein coupled receptor protein signaling pathway; insulin-like growth factor receptor signaling pathway; liver development; organ regeneration; positive regulation of apoptosis; post-embryonic development; receptor-mediated endocytosis; response to retinoic acid; signal transduction; spermatogenesis Disease: Hepatocellular Carcinoma |
| NCBI Summary: | This gene encodes a receptor for both insulin-like growth factor 2 and mannose 6-phosphate. The binding sites for each ligand are located on different segments of the protein. This receptor has various functions, including in the intracellular trafficking of lysosomal enzymes, the activation of transforming growth factor beta, and the degradation of insulin-like growth factor 2. Mutation or loss of heterozygosity of this gene has been association with risk of hepatocellular carcinoma. The orthologous mouse gene is imprinted and shows exclusive expression from the maternal allele; however, imprinting of the human gene may be polymorphic, as only a minority of individuals showed biased expression from the maternal allele (PMID:8267611). [provided by RefSeq, Nov 2015] |
| UniProt Code: | P11717 |
| NCBI GenInfo Identifier: | 317373416 |
| NCBI Gene ID: | 3482 |
| NCBI Accession: | P11717. 3 |
| UniProt Secondary Accession: | P11717,Q7Z7G9, Q96PT5, |
| UniProt Related Accession: | P11717 |
| Molecular Weight: | 274,375 Da |
| NCBI Full Name: | Cation-independent mannose-6-phosphate receptor |
| NCBI Synonym Full Names: | insulin like growth factor 2 receptor |
| NCBI Official Symbol: | IGF2R |
| NCBI Official Synonym Symbols: | MPR1; MPRI; CD222; CIMPR; M6P-R; MPR300; CI-M6PR; MPR 300; M6P/IGF2R |
| NCBI Protein Information: | cation-independent mannose-6-phosphate receptor |
| UniProt Protein Name: | Cation-independent mannose-6-phosphate receptor |
| UniProt Synonym Protein Names: | 300 kDa mannose 6-phosphate receptor; MPR 300; Insulin-like growth factor 2 receptor; Insulin-like growth factor II receptor; IGF-II receptor; M6P/IGF2 receptor; M6P/IGF2R; CD_antigen: CD222 |
| Protein Family: | Cation-independent mannose-6-phosphate receptor |
| UniProt Gene Name: | IGF2R |
| UniProt Entry Name: | MPRI_HUMAN |
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | RLU | Average | Corrected |
| 8000 | 64608 78270 | 71439 | 71418 |
| 4000 | 34687 36309 | 35498 | 35477 |
| 2000 | 17804 17304 | 17554 | 17533 |
| 1000 | 8066 9110 | 8588 | 8567 |
| 500 | 4179 4035 | 4107 | 4086 |
| 250 | 1945 1789 | 1867 | 1846 |
| 125.00 | 730 764 | 747 | 726 |
| 0 | 21 21 | 21 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IGF2R were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IGF2R were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 419.94 | 789.36 | 3128.14 | 412.07 | 809.34 | 3137.23 |
| Standard deviation | 43.38 | 94.72 | 253.07 | 43.47 | 82.80 | 209.88 |
| C V (%) | 10.33 | 12.00 | 8.09 | 10.55 | 10.23 | 6.69 |
Recovery
The recovery of Human IGF2R spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 95-108 | 101 |
| EDTA plasma (n=5) | 94-106 | 99 |
| Cell culture media (n=5) | 89-104 | 95 |
Linearity
Samples were spiked with high concentrations of Human IGF2R and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 100-113 | 89-100 | 90-105 |
| Average (%) | 106 | 95 | 96 | |
| 1:4 | Range (%) | 91-103 | 85-96 | 89-102 |
| Average (%) | 97 | 91 | 97 | |
| 1:8 | Range (%) | 88-103 | 88-104 | 84-96 |
| Average (%) | 95 | 95 | 91 | |
| 1:16 | Range (%) | 99-113 | 94-108 | 90-101 |
| Average (%) | 106 | 101 | 95 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro CLIA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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