New Product Focus: Lysozyme ELISA

New Product Focus
Lysozyme ELISA for the detection of human lysozyme in serum, plasma or stool.

The Calbiotech Lysozyme Assay Kit (Cat.# LS162C) provides researchers with a sensitive assay to measure levels of lysozyme in human samples. This direct sandwich ELISA assay can detect lysozyme from 50ng/ml down to 0.75 ng/mL.

Lysozyme is one of the anti-microbial agents present in spleen, lung, kidney, white blood cells, plasma, saliva, and tears and is also found in human milk. Lysozyme has 130 amino acids and its natural substrate is the bacterial cell wall peptidoglycan. Since synthesized by granulocytes and macrophages, lysozyme can act as a useful marker for myelomonocytic cells. Increased levels of lysozyme in urine and serum are diagnostic indicators for acute monocytic leukemia and acute myelomonycytic leukemia. Elevated lysozyme levels were found in synovial fluids of the inflammatory arthritides and osteoarthritis. Salivary lysozyme, a marker for oral infection and hyperglycemia, might display a significant relationship with hypertension, an early stage of cardiovascular disease.

Increased serum lysozyme activity is also present in tuberculosis, sarcoidosis, megaloblastic anemias, acute bacterial infections, ulcerative colitis and Crohns disease. Elevated lysozyme levels in urine and serum occur during severe renal insufficiency, renal transplant rejection, urinary tract infections, glomerulonephritis and nephrosis.

Principal of the Assay
The human Lysozyme ELISA kit is designed for detection of Lysozyme in human plasma, serum, or stool. This assay employs a quantitative sandwich enzyme immunoassay technique that measures Lysozyme in 75 minutes. A monoclonal antibody specific for Lysozyme has been pre-coated onto a microplate.

Lysozyme in standards and samples is sandwiched by the immobilized antibody and an HRP-conjugate monoclonal detection antibody specific for Lysozyme. All unbound material is then washed away and a peroxidase enzyme substrate is
added. The color development is stopped and the intensity of the color is measured.

Assay Specifications
Run time: 75min
Dynamic range: 0.75 – 50ng/ml
Sample type: Serum, Plasma, Stool
Sample volume: 25ul

This new product is easy to use, more accurate, and faster than its competitors and it comes with the great value and unmatched quality that are built in to all Calbiotech kits. To download the package insert, MSDS or to place an order for immediate delivery, please visit our website at www.calbiotech.com or CLICK HERE!

Preventing Contamination: Tweak your technique.

Calbiotech Immunoassay kits are easy to use, stable and reliable kits. Each kit comes with a package insert that is short and so vividly written that even a new ELISA user (technician) can comfortably perform the assay with a great accuracy and success. Nevertheless, like any other analytical measurement methods, ELISA is also susceptible to some contamination if we do not handle the reagents and/or instruments properly.

Today we will look at some common contamination which can occur during simple assay performance and ways to prevent them.

Cross-Contamination and Prevention

Sample-to-sample (carry-over)

In an assay involving multiple samples, remnants of sample A, after delivery, can mix with the subsequent sample B. This cross-contamination between samples may cause a pseudo positive test result.

Prevention:

• Change the tip after each sample.

Pipette-to-sample

Contaminated tips and/or a contaminated pipette can contaminate samples.

Prevention:

• Change the tip after each sample.
• Only use new or sterilized pipette tips.
• If you think the source of contamination is the pipette, clean or autoclave the parts of the pipette which are in contact with the sample.

Sample-to-pipette

Contamination can occur if sample or any other fluid is allowed to enter the cone of the pipette.

Prevention:

• Keeping the pipette vertical during pipetting can prevent liquids from running into the pipette body. Avoid inclining your pipette excessively.
• Always store the pipette vertically.
• Release the plunger slowly to prevent samples from jumping into the cone of the pipette.

Well-to-well

During assay performance, the washing step in the assay procedure is very critical. It is very possible that wells can cross-talk between each other. Make sure you consider the following points:

Prevention:

• Do not over fill the wells with wash buffer during the washing step.
• Clean your washing machine system (or bottle, if manual washing is used) regularly.
• Use clean distilled or de-ionized water while diluting the concentrate wash buffer provided in Calbiotech ELISA kits.

We hope you found our quick note on preventing cross contamination useful. Please share anything you think we missed in the comment section of this blog.

For technical support with running any of our assays, please contact support@calbiotech.com and we will get back to you immediately.