Mouse IL-4 ELISA Kit (MOES00662)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- IL4, BCGF-1, BCGF1, BSF-1, BSF1
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||31.25-2000 pg/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Mouse IL4 ELISA Kit in samples. No significant cross-reactivity or interference between Mouse IL4 ELISA Kit 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 IL4. 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 IL4 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 IL4, 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 IL4. The concentration of Mouse IL4 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||IL4: Participates in at least several B-cell activation processes as well as of other cell types. It is a costimulator of DNA-synthesis. It induces the expression of class II MHC molecules on resting B-cells. It enhances both secretion and cell surface expression of IgE and IgG1. It also regulates the expression of the low affinity Fc receptor for IgE (CD23) on both lymphocytes and monocytes. Genetic variations in IL4 may be a cause of susceptibility to ischemic stroke (ISCHSTR); also known as cerebrovascular accident or cerebral infarction. A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors. Belongs to the IL-4/IL-13 family. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Cell cycle regulation; Motility/polarity/chemotaxis; Secreted, signal peptide; Cytokine; Secreted
Cellular Component: extracellular space; extracellular region; external side of plasma membrane
Molecular Function:growth factor activity; hematopoietin/interferon-class (D200-domain) cytokine receptor binding; cytokine activity; interleukin-4 receptor binding
Biological Process: positive regulation of isotype switching to IgG isotypes; negative regulation of osteoclast differentiation; positive regulation of transcription, DNA-dependent; microglial cell activation; regulation of proton transport; positive regulation of activated T cell proliferation; positive regulation of interleukin-10 production; positive regulation of isotype switching to IgE isotypes; B cell costimulation; positive regulation of MHC class II biosynthetic process; positive regulation of cell proliferation; positive regulation of B cell proliferation; positive regulation of T cell proliferation; positive regulation of interleukin-13 production; negative regulation of macrophage activation; cholesterol metabolic process; negative regulation of chronic inflammatory response; regulation of immune response; B cell activation; negative regulation of nitric oxide biosynthetic process; negative regulation of acute inflammatory response; positive regulation of mast cell degranulation; defense response to protozoan; T-helper 1 cell lineage commitment; positive regulation of peptidyl-tyrosine phosphorylation; positive regulation of immunoglobulin production; positive regulation of chemokine biosynthetic process; positive regulation of T cell differentiation; innate immune response in mucosa; positive regulation of tyrosine phosphorylation of Stat5 protein; positive regulation of B cell activation; regulation of inflammatory response; positive regulation of transcription factor activity; immune response; positive regulation of transcription from RNA polymerase II promoter; positive regulation of protein amino acid phosphorylation; T-helper 2 cell differentiation; positive regulation of defense response to virus by host; negative regulation of transcription, DNA-dependent; negative regulation of T cell activation
|NCBI GenInfo Identifier:||124338|
|NCBI Gene ID:||16189|
|NCBI Accession:||P07750. 1|
|UniProt Related Accession:||P07750|
|Molecular Weight:||15,834 Da|
|NCBI Full Name:||Interleukin-4|
|NCBI Synonym Full Names:||interleukin 4|
|NCBI Official Symbol:||Il4|
|NCBI Official Synonym Symbols:||Il-4; BSF-1|
|NCBI Protein Information:||interleukin-4; IGG1 induction factor; B-cell growth factor 1; B-cell stimulatory factor 1; lymphocyte stimulatory factor 1; B-cell IgG differentiation factor|
|UniProt Protein Name:||Interleukin-4|
|UniProt Synonym Protein Names:||B-cell IgG differentiation factor; B-cell growth factor 1; B-cell stimulatory factor 1; BSF-1; IGG1 induction factor; Lymphocyte stimulatory factor 1|
|UniProt Gene Name:||Il4|
|UniProt Entry Name:||IL4_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 IL4 ELISA Kit were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse IL4 ELISA Kit were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.66||4.48||3.20||6.41||5.10||3.80|
The recovery of Mouse IL4 ELISA Kit 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)||95-111||101|
|Cell culture media (n=5)||92-108||98|
Samples were spiked with high concentrations of Mouse IL4 ELISA Kit 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.