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Rat 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1) ELISA Kit (RTEB0191)

SKU:
RTEB0191
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P10686
ELISA Type:
Sandwich
Reactivity:
Rat
€649
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Description

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Rat 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1) ELISA Kit

The Rat 1-Phosphatidylinositol-4,5-Bisphosphate Phosphodiesterase Gamma-1 (PLCG1) ELISA Kit is a reliable tool for the precise measurement of PLCG1 levels in rat samples, including serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers accurate and reproducible results, making it suitable for a variety of research applications.PLCG1 is a key enzyme involved in the regulation of intracellular signaling pathways, particularly in cell growth, differentiation, and immune response. Dysregulation of PLCG1 has been linked to various diseases, including cancer, autoimmune disorders, and neurological conditions.

Therefore, monitoring PLCG1 levels can provide valuable insights into the pathogenesis of these diseases and facilitate the development of targeted therapeutic strategies.Overall, the Rat PLCG1 ELISA Kit offers a powerful tool for researchers studying the role of PLCG1 in physiological and pathological processes, providing a comprehensive understanding of its functions and potential implications in disease mechanisms.

Product Name:Rat 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1) ELISA Kit
SKU:RTEB0191
Size:96T
Target:Rat 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-1 (Plcg1)
Synonyms:Phosphoinositide phospholipase C-gamma-1, Phospholipase C-gamma-1, PLC-gamma-1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:0.312-20ng/mL
Sensitivity:0.169ng/mL
Intra CV:Provided with the Kit
Inter CV:Provided with the Kit
Linearity:Provided with the Kit
Recovery:Provided with the Kit
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:P10686
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1
Sub Unit:Interacts (via SH2 domain) with FGFR1, FGFR2, FGFR3 and FGFR4 (phosphorylated). Interacts (via SH3 domain) with AGAP2. 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 (By similarity). Interacts with RALGPS1. Interacts (via the SH2 domains) with VIL1 (phosphorylated at C-terminus tyrosine phosphorylation sites). Interacts (via SH2 domain) with RET (By similarity). Interacts with FLT1 (tyrosine-phosphorylated). Interacts with AXL, FLT4 and KIT. Interacts (via SH2 domain) with PDGFRA and PDGFRB (tyrosine phosphorylated) (By similarity). Interacts with PIP5K1C (By similarity). Interacts with NTRK1 and NTRK2 (phosphorylated upon ligand-binding) (By similarity). Interacts with SYK; activates PLCG1 (By similarity). Interacts with TESPA1 (By similarity). Interacts with GRB2, LAT and THEMIS upon TCR activation in thymocytes.
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. Protein type: EC 3.1.4.11; Phospholipase; Motility/polarity/chemotaxis; Carbohydrate Metabolism - inositol phosphateCellular Component: signalosome; ruffle; neuron projection; cell projection; lamellipodium; cytoplasm; plasma membrane; intercellular junction; cytosolMolecular Function: glutamate receptor binding; signal transducer activity; protein binding; neurotrophin TRKA receptor binding; phosphoprotein binding; calcium ion binding; receptor tyrosine kinase binding; protein kinase binding; phosphoinositide phospholipase C activity; insulin receptor bindingBiological Process: epidermal growth factor receptor signaling pathway; response to gravity; cell migration; in utero embryonic development; phospholipid catabolic process; positive regulation of blood vessel endothelial cell migration; protein secretion; calcium-mediated signaling; response to morphine; signal transduction; response to organic nitrogen; T cell receptor signaling pathway; positive regulation of angiogenesis; response to hydrogen peroxide; inositol trisphosphate biosynthetic process; calcium ion transport; positive regulation of release of sequestered calcium ion into cytosol
UniProt Protein Details:
NCBI Summary:
UniProt Code:P10686
NCBI GenInfo Identifier:683466219
NCBI Gene ID:
NCBI Accession:KFZ60428.1
UniProt Secondary Accession:P10686
UniProt Related Accession:P10686
Molecular Weight:148,548 Da
NCBI Full Name:1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1, partial
NCBI Synonym Full Names:
NCBI Official Symbol:
NCBI Official Synonym Symbols:
NCBI Protein Information:
UniProt Protein Name:1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1
UniProt Synonym Protein Names:Phosphoinositide phospholipase C-gamma-1; Phospholipase C-gamma-1; PLC-gamma-1
Protein Family:1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase
UniProt Gene Name:Plcg1
UniProt Entry Name:PLCG1_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.

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