Description
Mouse 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1) ELISA Kit
The Mouse 1-Phosphatidylinositol-4,5-Bisphosphate Phosphodiesterase Gamma 1 (PLCG1) ELISA Kit is a reliable tool for the precise measurement of PLCG1 levels in mouse serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring accurate and consistent results for a variety of research applications. PLCG1 is a key enzyme involved in intracellular signaling pathways, particularly in the regulation of cell growth, differentiation, and immune responses.
Dysregulation of PLCG1 has been implicated in various diseases including cancer, immune disorders, and neurological conditions, making it a valuable biomarker for disease research and therapeutic development. By utilizing the Mouse 1-Phosphatidylinositol-4,5-Bisphosphate Phosphodiesterase Gamma 1 (PLCG1) ELISA Kit, researchers can gain valuable insights into the role of PLCG1 in disease pathogenesis and explore potential therapeutic targets for treatment.
| Product Name: | Mouse 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1) ELISA Kit |
| SKU: | MOEB0237 |
| Size: | 96T |
| Target: | Mouse 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1) |
| Synonyms: | Phosphoinositide phospholipase C-gamma-1, Phospholipase C-gamma-1, PLC-gamma-1, Plcg-1 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Mouse |
| Detection Range: | 0.156-10ng/ml |
| Sensitivity: | 0.084ng/mL |
| Intra CV: | 6.1% | ||||||||||||||||||||
| Inter CV: | 8.3% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Mediates the production of the second messenger molecules diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3). Plays an important role in the regulation of intracellular signaling cascades. Becomes activated in response to ligand-mediated activation of receptor-type tyrosine kinases, such as PDGFRA, PDGFRB, FGFR1, FGFR2, FGFR3 and FGFR4. Plays a role in actin reorganization and cell migration. |
| Uniprot: | Q62077 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant mouse 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1 |
| Sub Unit: | Interacts (via SH2 domain) with FGFR1, FGFR2, FGFR3 and FGFR4 (phosphorylated). Interacts with RALGPS1. Interacts (via SH2 domains) with VIL1 (phosphorylated at C-terminus tyrosine phosphorylation sites). Interacts (via SH2 domain) with RET (By similarity). Interacts with AGAP2 via its SH3 domain. Interacts with LAT (phosphorylated) upon TCR activation. Interacts (via SH3 domain) with the Pro-rich domain of TNK1. Associates with BLNK, VAV1, GRB2 and NCK1 in a B-cell antigen receptor-dependent fashion. Interacts with CBLB in activated T-cells; which inhibits phosphorylation. Interacts with SHB. Interacts (via SH3 domain) with the Arg/Gly-rich-flanked Pro-rich domains of KHDRBS1/SAM68. This interaction is selectively regulated by arginine methylation of KHDRBS1/SAM68. Interacts with INPP5D/SHIP1, THEMIS and CLNK. Interacts with FLT4 and KIT. Interacts with AXL (By similarity). Interacts with SYK; activates PLCG1 (By similarity). Interacts with FLT1 (tyrosine-phosphorylated). Interacts (via SH2 domain) with PDGFRA and PDGFRB (tyrosine phosphorylated). Interacts with PIP5K1C. Interacts with NTRK1 and NTRK2 (phosphorylated upon ligand-binding). Interacts with TESPA1 (By similarity). Interacts with GRB2, LAT and THEMIS upon TCR activation in thymocytes; the association is weaker in the absence of TESPA1. |
| Research Area: | Cardiovascular |
| Subcellular Location: | Cell projection Lamellipodium Cell projection Ruffle Rapidly redistributed to ruffles and lamellipodia structures in response to epidermal growth factor (EGF) treatment. |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | PLCG1: a calcium dependent phosphatidylinositol-specific phospholipase C. The activated enzyme produces the second messenger molecules diacylglycerol and inositol 1,4,5-trisphosphate. Phosphorylated and activated by tyrosine kinases in response to signaling through a variety of growth factor receptors and immune system receptors. |
| UniProt Protein Details: | Protein type:Carbohydrate Metabolism - inositol phosphate; Motility/polarity/chemotaxis; Phospholipase; EC 3.1.4.11 Cellular Component: signalosome; ruffle; neuron projection; cell projection; lamellipodium; cytoplasm; plasma membrane; intercellular junction; cytosol Molecular Function:glutamate receptor binding; phosphoric diester hydrolase activity; hydrolase activity; metal ion binding; neurotrophin TRKA receptor binding; phosphoprotein binding; calcium ion binding; receptor tyrosine kinase binding; phosphoinositide phospholipase C activity; protein kinase binding; insulin receptor binding; protein binding; signal transducer activity Biological Process: epidermal growth factor receptor signaling pathway; cell migration; positive regulation of angiogenesis; phospholipid catabolic process; in utero embryonic development; inositol trisphosphate biosynthetic process; calcium ion transport; positive regulation of blood vessel endothelial cell migration; calcium-mediated signaling; protein secretion; lipid metabolic process; positive regulation of release of sequestered calcium ion into cytosol; signal transduction; T cell receptor signaling pathway; lipid catabolic process |
| UniProt Code: | Q62077 |
| NCBI GenInfo Identifier: | 62635484 |
| NCBI Gene ID: | 18803 |
| NCBI Accession: | AAX90611.1 |
| UniProt Secondary Accession: | Q62077,Q6P1G1, |
| UniProt Related Accession: | Q62077 |
| Molecular Weight: | 149,668 Da |
| NCBI Full Name: | Plcg1 |
| NCBI Synonym Full Names: | phospholipase C, gamma 1 |
| NCBI Official Symbol: | Plcg1 |
| NCBI Official Synonym Symbols: | Cded; Plc-1; Plcg-1; AI894140; Plc-gamma1 |
| NCBI Protein Information: | 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1; PLC-gamma-1; phospholipase C-gamma-1; phosphoinositide phospholipase C-gamma-1; cell differentiation and embryonic development; 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma 1; 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 |
| UniProt Protein Name: | 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1 |
| UniProt Synonym Protein Names: | Phosphoinositide phospholipase C-gamma-1; Phospholipase C-gamma-1 |
| Protein Family: | 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase |
| UniProt Gene Name: | Plcg1 |
| UniProt Entry Name: | PLCG1_MOUSE |
| 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. |
