Human IL-4 ELISA Kit (HUES01387)
- 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 Human IL4 ELISA Kit in samples. No significant cross-reactivity or interference between Human 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 Human 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 Human 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 Human 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 Human IL4. The concentration of Human 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; Cytokine; Motility/polarity/chemotaxis; Secreted; Secreted, signal peptide
Chromosomal Location of Human Ortholog: 5q31. 1
Cellular Component: extracellular space; external side of plasma membrane
Molecular Function:protein binding; growth factor activity; cytokine activity; interleukin-4 receptor binding
Biological Process: regulation of isotype switching; 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; female pregnancy; positive regulation of activated T cell proliferation; chemotaxis; response to organic cyclic substance; positive regulation of isotype switching to IgE isotypes; positive regulation of interleukin-10 production; B cell costimulation; connective tissue growth factor biosynthetic process; positive regulation of MHC class II biosynthetic process; positive regulation of B cell proliferation; positive regulation of T cell proliferation; positive regulation of interleukin-13 production; negative regulation of macrophage activation; response to nutrient; response to drug; cholesterol metabolic process; regulation of immune response; negative regulation of chronic inflammatory response; T-helper 2 type immune response; regulation of phosphorylation; B cell activation; negative regulation of nitric oxide biosynthetic process; negative regulation of acute inflammatory response; defense response to protozoan; T-helper 1 cell lineage commitment; response to ethanol; positive regulation of chemokine biosynthetic process; positive regulation of tyrosine phosphorylation of Stat5 protein; positive regulation of T cell differentiation; innate immune response in mucosa; retina development in camera-type eye; B cell differentiation; response to cytokine stimulus; cellular defense response; immune response; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription factor activity; T-helper 2 cell differentiation; negative regulation of transcription, DNA-dependent; positive regulation of defense response to virus by host; negative regulation of apoptosis
Disease: Stroke, Ischemic
|NCBI Summary:||The protein encoded by this gene is a pleiotropic cytokine produced by activated T cells. This cytokine is a ligand for interleukin 4 receptor. The interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. STAT6, a signal transducer and activator of transcription, has been shown to play a central role in mediating the immune regulatory signal of this cytokine. This gene, IL3, IL5, IL13, and CSF2 form a cytokine gene cluster on chromosome 5q, with this gene particularly close to IL13. This gene, IL13 and IL5 are found to be regulated coordinately by several long-range regulatory elements in an over 120 kilobase range on the chromosome. Two alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||124337|
|NCBI Gene ID:||3565|
|NCBI Accession:||P05112. 1|
|UniProt Secondary Accession:||P05112,Q14630, Q6NZ77,|
|UniProt Related Accession:||P05112|
|Molecular Weight:||15,797 Da|
|NCBI Full Name:||Interleukin-4|
|NCBI Synonym Full Names:||interleukin 4|
|NCBI Official Symbol:||IL4|
|NCBI Official Synonym Symbols:||BSF1; IL-4; BCGF1; BSF-1; BCGF-1|
|NCBI Protein Information:||interleukin-4; binetrakin; pitrakinra; B cell growth factor 1; interleukin 4 variant 2; B_cell stimulatory factor 1; lymphocyte stimulatory factor 1|
|UniProt Protein Name:||Interleukin-4|
|UniProt Synonym Protein Names:||B-cell stimulatory factor 1; BSF-1; Binetrakin; Lymphocyte stimulatory factor 1; Pitrakinra|
|UniProt Gene Name:||IL4|
|UniProt Entry Name:||IL4_HUMAN|
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 Human 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 Human IL4 ELISA Kit were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.03||4.22||3.50||6.13||5.01||3.00|
The recovery of Human 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)||93-108||98|
|Cell culture media (n=5)||90-104||96|
Samples were spiked with high concentrations of Human 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 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 90 µL 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.