Mouse PLCg1 (Phospholipase C Gamma 1) ELISA Kit (MOES01358)
- Product type:
- ELISA Kit
- ELISA Type:
|Detection Range:||1.56-100 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Mouse PLCg1 in samples. No significant cross-reactivity or interference between Mouse PLCg1 and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse PLCg1. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse PLCg1 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 Mouse PLCg1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse PLCg1. The concentration of Mouse PLCg1 in samples can be calculated by comparing the OD of the samples to the standard curve.
|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
|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|
As the OD 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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse PLCg1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse PLCg1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.66||4.69||3.08||5.42||5.18||3.53|
The recovery of Mouse PLCg1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||90-104||95|
|Cell culture media (n=5)||84-98||90|
Samples were spiked with high concentrations of Mouse PLCg1 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)|
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.
|Micro ELISA 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||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.