ELISA, an acronym for Enzyme-Linked Immunosorbent Assay, is an established technology used in detecting the presence of an antigen in a given biological sample. In detecting the target antigens, ELISA relies on antibodies by making use of specific antibody antigens interactions.
In the ELISA principles, the antigens are immobilized to a solid-state directly or through the captured antibody itself that is immobilized on the surface. In streamlining immunodetection experiments, there are target-specific ELISA kits from various manufacturers which can be used. This ELISA kit targets the human IL4 ELISA kit Picokine.
Human IL-4 ELISA Kit
The Human interleukin 4 ELISA kit, otherwise known as Hu Il-4, quantitates human Il 4 in cell culture supernatants, cell lysates, serum, plasma, and buffered solution.
Target information of human IL-4 ELISA Kit Picokine
Human IL-4 ELISA kit is produced by activated T cells, which is a pleiotropic and immune-modulatory cytokine. This interleukin Multiples has a ligand for the IL 4 receptor and has the broadest range of actions. The receptor of the IL-4 binds to IL13, which may result in overlapping functions of cytokine and IL13 The Signal transducer and activator of transcription, which is known as STAT6, plays a crucial role in the meditating process of the immune regulatory signal of IL-4. The protein that is in the IL-4 has a predicted amino acid of 153 as its length, 17.5KDa as mass, and a secreted subcellular localization.
Principle of the human IL-4 kit method
The Design of the human IL-4 kit measures the amount of the target bound which exists between a matched antibody pair. In the wells of the supplied microplate, target-antibody is pre-coated in it. After that, samples, standards, or controls are added into the wells which bind the immobilized antibody. Through the addition of the second antibody, a sandwich is then formed. After the formation, a substrate solution is added, which reacts with the enzyme antibody target, producing measurable signals.
Storage and handling of the human IL-4 ELISA kit
In storing the Il-4 kit, proper care needs to be given to it to avoid damage. In storage, it is best stored at 4oC for six months. In storing for twelve months, the storage should be at -20o C. Multiple freeze-thaw cycles should be avoided in keeping the kit in its accurate state for use. If the kit is transported from one place to another, it is best shipped with a block of wet ice.
Kit components of human IL-4 ELISA
There are different components for this kit, and each has its catalog number, description, and quantity.
EK0404-CAP is an anti-human IL-4 pre-coated 96-well striped microplate and is of a single quantity.
EK0404-ST is the Human IL-4 standard. Its quality is two vials, 10ng/tube.
EK0404-DA is a Human IL4 Biotinylated antibody (100x) of quantity 100ul.
AR1103 is an Avidin-Biotin-Peroxidase Complex (100x), and its quantity is 100ul
AR1106-1 is a Sample Diluent of quantity 30ml
AR1106-2 is an Antibody Diluent that has a quantity of 12ml.
AR1106-3 is an Avidin-Biotin-Peroxidase Diluent of 12ml in quantity.
AR1104 is a Color Developing Reagent (TMB) that has 10ml as its quality.
AR1105 is a Stop Solution of 10ml in quality.
AR1106-5 is a Wash Buffer (25x) of 20ml in quality.
PLA-SEA is an Adhesive plate sealer that is 4 in quality.
Advantages of ELISA
ELISA has a high sensitivity to detect antigens at the level of a picogram, which is very specific. And this is a result of the antibodies used.
Protocols in ELISA are easy to follow and do not require difficulties.
ELISA is quantitative, which means the concentrations of the antigens in biological samples can be determined.
It can test different samples like the serum, plasma, cellular and tissue extracts, saliva, urine, etc.
It has a high throughput which means that the commercial ELISA kits are available in 96 well plate format.
Disadvantages of ELISA
The readout must be obtained quickly because the detection is based on enzyme and substrate reactions.
ELISA has limited information in proportion to the amount or presence of the antigens in the biological samples.
ELISA Analysis
In analyzing ELISA data, choosing the most appropriate method for the sample gives a better result. ELISA assays can be classified as qualitative ELISA, semi-qualitative ELISA, and quantitative ELSA. Qualitative ELISA is used in determining if the antigens are represented in a sample or not. Semi-qualitative ELISA is meant for the relative comparison of antigens in the samples. While for quantitative ELISA, it calculates the number of antigens that are represented in the sample.
Conclusion
The human IL-4 ELISA kit Picokine is of quality that has been tested through its sensitivity, specificity, precision, and various consistency. The kit’s sensitivity is 1.5pg/ml with an assay range of 15.5pg/ml to 1000pg/ml.