Description
Rat Substance-P receptor (Tacr1) ELISA Kit
The Rat Substance P Receptor (TACR1) ELISA Kit is a powerful tool for accurate measurement of TACR1 levels in rat samples, including serum, plasma, and tissue homogenates. With exceptional sensitivity and specificity, this kit delivers precise and reproducible results, making it suitable for various research applications.TACR1, also known as the Substance P receptor, plays a crucial role in mediating pain perception and neurogenic inflammation. Its dysregulation has been linked to conditions such as chronic pain, inflammatory diseases, and psychiatric disorders, making it a valuable biomarker for studying these conditions and exploring potential therapeutic interventions.
With the Rat Substance P Receptor (TACR1) ELISA Kit, researchers can confidently investigate the role of TACR1 in various physiological and pathological processes, advancing our understanding of this important receptor and its implications for health and disease.
| Product Name: | Rat Substance-P receptor (Tacr1) ELISA Kit |
| SKU: | RTEB0257 |
| Size: | 96T |
| Target: | Rat Substance-P receptor (Tacr1) |
| Synonyms: | NK-1 receptor, Tachykinin receptor 1, NK-1R, SPR, Tac1r |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Rat |
| Detection Range: | 0.312-20ng/mL |
| Sensitivity: | 0.176ng/mL |
| Intra CV: | Provided with the Kit |
| Inter CV: | Provided with the Kit |
| Linearity: | Provided with the Kit |
| Recovery: | Provided with the Kit |
| Function: | This is a receptor for the tachykinin neuropeptide substance P. It is probably associated with G proteins that activate a phosphatidylinositol-calcium second messenger system. The rank order of affinity of this receptor to tachykinins is: substance P > substance K > neuromedin-K. |
| Uniprot: | P14600 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant rat Substance-P receptor |
| Sub Unit: | Interacts with ARRB1. |
| Subcellular Location: | Cell membrane Multi-pass membrane protein |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | TACR1: This is a receptor for the tachykinin neuropeptide substance P. It is probably associated with G proteins that activate a phosphatidylinositol-calcium second messenger system. The rank order of affinity of this receptor to tachykinins is: substance P > substance K > neuromedin-K. Belongs to the G-protein coupled receptor 1 family. |
| UniProt Protein Details: | Protein type:Membrane protein, multi-pass; Receptor, GPCR; GPCR, family 1; Membrane protein, integral Cellular Component: cell surface; cytoplasm; cytosol; dendrite; integral to plasma membrane; plasma membrane Molecular Function:substance P receptor activity Biological Process: acute inflammatory response; angiotensin mediated drinking behavior; associative learning; behavioral response to pain; cell surface receptor linked signal transduction; eating behavior; elevation of cytosolic calcium ion concentration; learning and/or memory; long-term memory; neuropeptide signaling pathway; operant conditioning; positive regulation of action potential; positive regulation of blood pressure; positive regulation of epithelial cell proliferation; positive regulation of hormone secretion; positive regulation of leukocyte migration; positive regulation of lymphocyte proliferation; positive regulation of ossification; positive regulation of saliva secretion; positive regulation of stress fiber formation; positive regulation of synaptic transmission, cholinergic; positive regulation of synaptic transmission, GABAergic; positive regulation of vascular permeability; positive regulation of vasoconstriction; regulation of blood pressure; regulation of smooth muscle cell migration; regulation of smooth muscle cell proliferation; response to electrical stimulus; response to estradiol stimulus; response to ethanol; response to heat; response to hormone stimulus; response to morphine; response to nicotine; response to organic cyclic substance; response to ozone; response to pain; response to progesterone stimulus; sensory perception of pain; smooth muscle contraction involved in micturition; sperm ejaculation; tachykinin signaling pathway |
| NCBI Summary: | mediates neuropeptide signaling; may play a role in pain sensitivity; may mediate baroreflex transmission for blood pressure regulation [RGD, Feb 2006] |
| UniProt Code: | P14600 |
| NCBI GenInfo Identifier: | 128360 |
| NCBI Gene ID: | 24807 |
| NCBI Accession: | P14600.1 |
| UniProt Related Accession: | P14600 |
| Molecular Weight: | 46,366 Da |
| NCBI Full Name: | Substance-P receptor |
| NCBI Synonym Full Names: | tachykinin receptor 1 |
| NCBI Official Symbol: | Tacr1 |
| NCBI Official Synonym Symbols: | Tac1r |
| NCBI Protein Information: | substance-P receptor |
| UniProt Protein Name: | Substance-P receptor |
| UniProt Synonym Protein Names: | NK-1 receptor; NK-1R; Tachykinin receptor 1 |
| UniProt Gene Name: | Tacr1 |
| UniProt Entry Name: | NK1R_RAT |
| 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. |
