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Datasheet MSDS Protocol

Porcine uPA ELISA Kit

Quantity
Quantity
$495.00
  • Catalog # PU0019
  • Detection Range 0, 50, 100, 250, 500, 1000pg/ml
  • Sensitivity 1.0pg/ml
  • Size 96 T
  • Assay Time 90 minutes
  • Sample 100 ul
  • Conjugate HRP
  • Instrument Microlate Reader
  • Precautions Competitive ELISA
  • SPECIFICATION
  • BACKGROUND
  • DOCUMENTS
  • REVIEW
  • FAQ

ADDITIONAL INFORMATION

PU0019

Porcine uPA ELISA Kit

Product Name

Porcine Urokinase plasminogen activator ELISA Kit

SAMPLE TYPE

100 ul

PRODUCT OVERVIEW

The ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of samples in serum, plasma, cell culture supernatants and urine.

INTENDED USE

This Porcine uPA ELISA Kit is intended for laboratory research use only and not for use in diagnostic or therapeutic procedures. The stop solution changes color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of Porcine uPA in the sample, this Porcine uPA ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of optical density versus Porcine uPA concentration. The concentration of the samples is then determined by comparing the O.D. of the samples to the standard curve.

STORAGE INSTRUCTIONS

Store at 4℃

COMPONENTS

Microtiter Plate
Enzyme Conjugate
Standards
Substrate A
Substrate B
Stop Solution
Wash Solution
Balance Solution

SAFETY NOTES

1. This kit contains materials with small quantities of sodium azide. Sodium azide reacts with lead and copper plumbing to form explosive metal azides. Upon disposal, flush drains with a large volume of water to prevent azide accumulation. Avoid ingestion and contact with eyes, skin or mucous membranes. In the case of contact, rinse the affected area with plenty of water. Observe all federal, state and local regulations for disposal.

2. All blood components and biological materials should be handled as potentially hazardous. Follow universal precautions as established by the Centers for Disease Control and Prevention and by the Occupational Safety and Health Administration when handling and disposing of infectious agents.

BACKGROUND

As an analytic biochemistry assay, ELISA involves detection of an 'analyte' (i.e. the specific substance whose presence is being quantitatively or qualitatively analyzed) in a liquid sample by a method that continues to use liquid reagents during the 'analysis' (i.e. controlled sequence of biochemical reactions that will generate a signal which can be easily quantified and interpreted as a measure of the amount of analyte in the sample) that stays liquid and remains inside a reaction chamber or well needed to keep the reactants contained.

As a heterogenous assay, ELISA separates some component of the analytical reaction mixture by adsorbing certain components onto a solid phase which is physically immobilized. In ELISA, a liquid sample is added onto a stationary solid phase with special binding properties and is followed by multiple liquid reagents that are sequentially added, incubated and washed followed by some optical change (e.g. color development by the product of an enzymatic reaction) in the final liquid in the well from which the quantity of the analyte is measured. The qualitative 'reading' usually based on detection of intensity of transmitted light by spectrophotometry, which involves quantitation of transmission of some specific wavelength of light through the liquid (as well as the transparent bottom of the well in the multiple-well plate format). The sensitivity of detection depends on amplification of the signal during the analytic reactions. Since enzyme reactions are very well known amplification processes, the signal is generated by enzymes which are linked to the detection reagents in fixed proportions to allow accurate quantification - thus the name 'enzyme linked'.

The analyte is also called the ligand because it will specifically bind or ligate to a detection reagent, thus ELISA falls under the bigger category of ligand binding assays. The ligand-specific binding reagent is 'immobilized', i.e., usually coated and dried onto the transparent bottom and sometimes also side wall of a well (the stationary 'solid phase 'solid substrate' here as opposed to solid microparticle/beads that can be washed away), which is usually constructed as a multiple-well plate known as the 'ELISA plate'. Conventionally, like other forms of immunoassays, the specificity of antigen-antibody type reaction is used because it is easy to raise an antibody specifically against an antigen in bulk as a reagent. Alternatively, if the analyte itself is an antibody, its target antigen can be used as the binding reagent.

DOCUMENTS

Datasheet

MSDS

Protocol

REVIEWS

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ELISA Kit FAQ

Your Guide to Understanding and Using ELISA Kits

A Competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is a type of immunoassay used to measure the concentration of an antigen in a sample. In this assay, the antigen in the sample competes with a labeled antigen for binding to a limited number of antibody binding sites. The signal produced is inversely proportional to the amount of antigen in the sample. Competitive ELISA is particularly useful for detecting small molecules and compounds, where direct or sandwich ELISAs might not be effective.
A Sandwich ELISA kit is designed to measure the amount of antigen in a sample by "sandwiching" the antigen between two layers of antibodies. The process starts with an antibody coated on the plate, which captures the antigen from the sample. A second antibody, linked to an enzyme, is then added to bind to the captured antigen. The enzyme-substrate reaction produces a measurable signal, directly proportional to the antigen concentration in the sample.

There are key differences between Sandwich and Competitive ELISA:

  • Principle: In Sandwich ELISA, the antigen is captured between two layers of antibodies. In Competitive ELISA, the antigen in the sample competes with a labeled antigen for binding to a limited number of antibody sites.
  • Signal Correlation: In Sandwich ELISA, the signal produced is directly proportional to the antigen concentration. In Competitive ELISA, the signal is inversely proportional to the antigen concentration.
  • Application: Sandwich ELISA is often used for detecting proteins and larger molecules, while Competitive ELISA is better suited for small molecules and haptens.

To ensure optimal performance and accuracy of your ELISA kit, proper storage is essential. Please adhere to the following guidelines:

  • Sealed ELISA Kits: Store the sealed kit at 2-8°C for a maximum of 6 months to preserve its integrity.
  • Opened ELISA Kits: Once the kit is opened, it should be stored at 2-8°C and used within one month, provided the components are not contaminated.
Sample Type Balance Solution Required
Cell Culture Supernatant Yes
Body Fluid Yes
Tissue Homogenate Yes
Undiluted Blood or undiluted serum No
Other Samples Please contact info@neoscientific.com for confirmation

For optimal results, the sample buffer should be Phosphate Buffered Saline (PBS). Please note that PBS is not provided in the kit, so you will need to prepare or obtain it separately. PBS helps in maintaining the pH and ionic strength of the samples, which is essential for the assay's accuracy and reproducibility.

We hope this Q&A provides a comprehensive overview of ELISA kits and assists you in your research or diagnostic applications. If you have any further questions, please do not hesitate to contact us.

Yes, running standards and samples in duplicate is highly recommended. This practice helps monitor assay precision and enhances the confidence in the results you obtain from the assay.
No, each kit uses a strip well microplate. This allows the user to analyze different numbers of samples at different times.
Each kit comes with a manual containing a graph of typical data obtained. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.
The kit has been validated for the sample types listed in the kit insert. Sample types other than those validated have not been tested. Contact Technical Service for further information.
It is recommended that only sample values that fall within the range of the standard curve be used. Values outside the range of the standard curve are generally non-linear, which can lead to incorrectly extrapolated values. Samples that generate values higher than the highest standard should be further diluted and the assay repeated. If samples fall below the range of the assay, the sample is considered to be non-detectable.

A blank is required for the calculations, as it reflects any subtle but significant performance changes from day to day and assay to assay. However, if the operator is experienced, there is no big need to run the blank.

For any further questions, please contact info@neoscientific.com for confirmation.

It is not recommended that you alter the volumes since our kits are designed for optimal performance at the given volumes.
Each kit contains components which have specific lot numbers to ensure that all of the components are performing optimally alone, as well as with all of the other components in the kit. QC testing is performed on these specific lots. It is never recommended to use your own components or components from other kits or vendors.
The sample may not contain the analyte. A matrix effect may be masking the detection. Over-dilution may cause the sample to fall below the range of the standard curve.
The optical density is affected by a number of physical conditions such as time and temperature. We suggest that you shorten or lengthen the final incubation with substrate solution to compensate.

There are several reasons for high background in your assay results:

  • Improper Washing: Check the volume of the washing buffer reservoir and make sure all recommended washing steps are performed correctly.
  • Substrate exposed to light: Exposure to light may result in a blue color of the substrate. To avoid this, keep solutions in the dark (vial) until they are ready to be dispensed into the plate.
  • Wrong Incubation Times/Temperatures: Generally, follow the test protocol regarding incubation times and temperatures. However, if all wells are intensely and equally colored with no intensity gradient observed in the standard dilution series, it may be necessary to observe the substrate reaction as the color is developing and stop the reaction sooner.
For additional assistance, please contact us at info@neoscientific.com