Description
Human Insulin-like growth factor 1 receptor (IGF1R) ELISA Kit
The Human Insulin-like Growth Factor 1 Receptor (IGF1R) ELISA Kit is specially designed for the precise measurement of IGF1R levels in human serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results, making it perfect for various research applications.IGF1R is a key receptor involved in mediating the effects of insulin-like growth factors, regulating cell growth, differentiation, and survival.
It plays a vital role in multiple physiological processes and is implicated in various diseases, such as cancer, diabetes, and aging-related conditions. Therefore, the Human IGF1R ELISA Kit is an essential tool for studying IGF1R biology and its potential therapeutic implications.
| Product Name: | Human Insulin-like growth factor 1 receptor (IGF1R) ELISA Kit |
| SKU: | HUEB1244 |
| Size: | 96T |
| Target: | Human Insulin-like growth factor 1 receptor (IGF1R) |
| Synonyms: | Insulin-like growth factor I receptor, IGF-I receptor, CD221 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Human |
| Detection Range: | 0.312-20ng/mL |
| Sensitivity: | 0.098ng/mL |
| Intra CV: | 5.2% | ||||||||||||||||||||
| Inter CV: | 8.0% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | When present in a hybrid receptor with INSR, binds IGF1. PubMed:12138094 shows that hybrid receptors composed of IGF1R and INSR isoform Long are activated with a high affinity by IGF1, with low affinity by IGF2 and not significantly activated by insulin, and that hybrid receptors composed of IGF1R and INSR isoform Short are activated by IGF1, IGF2 and insulin. In contrast, PubMed:16831875 shows that hybrid receptors composed of IGF1R and INSR isoform Long and hybrid receptors composed of IGF1R and INSR isoform Short have similar binding characteristics, both bind IGF1 and have a low affinity for insulin. |
| Uniprot: | P08069 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant human Insulin-like growth factor 1 receptor |
| Sub Unit: | Tetramer of 2 alpha and 2 beta chains linked by disulfide bonds. The alpha chains contribute to the formation of the ligand-binding domain, while the beta chain carries the kinase domain. Interacts with PIK3R1 and with the PTB/PID domains of IRS1 and SHC1 in vitro when autophosphorylated on tyrosine residues. Forms a hybrid receptor with INSR, the hybrid is a tetramer consisting of 1 alpha chain and 1 beta chain of INSR and 1 alpha chain and 1 beta chain of IGF1R. Interacts with ARRB1 and ARRB2. Interacts with GRB10. Interacts with RACK1. Interacts with SOCS1, SOCS2 and SOCS3. Interacts with 14-3-3 proteins. Interacts with NMD2. Interacts with MAP3K5. Interacts with STAT3. Found in a ternary complex with IGF1 and ITGAV:ITGB3 or ITGA6:ITGB4 (PubMed:19578119, PubMed:22351760). |
| Research Area: | Cancer |
| Subcellular Location: | Cell membrane Single-pass type I membrane protein |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | IGF1R: a receptor tyrosine kinase that binds insulin-like growth factor 1 (IGF1) with a high affinity and IGF2 with a lower affinity. Functions as an anti-apoptotic agent by enhancing cell survival. Can play a critical role in transformation events. Cleavage of the precursor generates alpha and beta subunits. Tetramer of 2 alpha and 2 beta chains linked by disulfide bonds. The alpha chains contribute to the formation of the ligand- binding domain, while the beta chain carries the kinase domain. Interacts with PIK3R1 and with the PTB/PID domains of IRS1 and SHC1 in vitro when autophosphorylated on tyrosine residues. Mutated in rare cases of pre- and post-natal growth retardation. One SNP associated with increased human longevity. Increased expression of IGF1R and other pathway members associated with progression and malignancy in a range of cancers. Inhibitors: AG1024, AEW541. |
| UniProt Protein Details: | Protein type:Protein kinase, tyrosine (receptor); Protein kinase, TK; Kinase, protein; Membrane protein, integral; EC 2.7.10.1; TK group; InsR family Chromosomal Location of Human Ortholog: 15q26.3 Cellular Component: integral to plasma membrane; intracellular membrane-bound organelle; membrane; plasma membrane; receptor complex Molecular Function:ATP binding; identical protein binding; insulin binding; insulin receptor binding; insulin receptor substrate binding; insulin-like growth factor binding; insulin-like growth factor I binding; insulin-like growth factor receptor activity; phosphoinositide 3-kinase binding; protein binding; protein-tyrosine kinase activity Biological Process: brain development; epidermis development; exocrine pancreas development; immune response; inactivation of MAPKK activity; insulin receptor signaling pathway; insulin-like growth factor receptor signaling pathway; male sex determination; mammary gland development; negative regulation of apoptosis; negative regulation of protein kinase B signaling cascade; negative regulation of transcription factor activity; phosphoinositide 3-kinase cascade; phosphoinositide-mediated signaling; positive regulation of cell migration; positive regulation of cell proliferation; positive regulation of DNA replication; positive regulation of MAPKKK cascade; positive regulation of mitosis; positive regulation of protein kinase B signaling cascade; protein amino acid autophosphorylation; protein tetramerization; regulation of JNK cascade; signal transduction Disease: Insulin-like Growth Factor I, Resistance To |
| NCBI Summary: | This receptor binds insulin-like growth factor with a high affinity. It has tyrosine kinase activity. The insulin-like growth factor I receptor plays a critical role in transformation events. Cleavage of the precursor generates alpha and beta subunits. It is highly overexpressed in most malignant tissues where it functions as an anti-apoptotic agent by enhancing cell survival. Alternatively spliced transcript variants encoding distinct isoforms have been found for this gene. [provided by RefSeq, May 2014] |
| UniProt Code: | P08069 |
| NCBI GenInfo Identifier: | 124240 |
| NCBI Gene ID: | 3480 |
| NCBI Accession: | P08069.1 |
| UniProt Secondary Accession: | P08069,Q14CV2, Q9UCC0, B1B5Y2, |
| UniProt Related Accession: | P08069 |
| Molecular Weight: | |
| NCBI Full Name: | Insulin-like growth factor 1 receptor |
| NCBI Synonym Full Names: | insulin like growth factor 1 receptor |
| NCBI Official Symbol: | IGF1R |
| NCBI Official Synonym Symbols: | IGFR; CD221; IGFIR; JTK13 |
| NCBI Protein Information: | insulin-like growth factor 1 receptor |
| UniProt Protein Name: | Insulin-like growth factor 1 receptor |
| UniProt Synonym Protein Names: | Insulin-like growth factor I receptor; IGF-I receptor |
| Protein Family: | Insulin-like growth factor 1 receptor |
| UniProt Gene Name: | IGF1R |
| UniProt Entry Name: | IGF1R_HUMAN |
| Component | Quantity (96 Assays) | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | -20°C |
| Lyophilized Standard | 2 | -20°C |
| Sample Diluent | 20ml | -20°C |
| Assay Diluent A | 10mL | -20°C |
| Assay Diluent B | 10mL | -20°C |
| Detection Reagent A | 120µL | -20°C |
| Detection Reagent B | 120µL | -20°C |
| Wash Buffer | 30mL | 4°C |
| Substrate | 10mL | 4°C |
| Stop Solution | 10mL | 4°C |
| Plate Sealer | 5 | - |
Other materials and equipment required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.
| Step | |
| 1. | Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C. |
| 2. | Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters. |
| 9. | After experiment, store all reagents according to the specified storage temperature respectively until their expiry. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
