Chromogenic LAL Endotoxin Assay Kit (160T)
$649.00
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Cat. No.: CLALE-160 (for 160T)
Description
Chromogenic LAL Endotoxin Assay Kit is a commonly used reagent kit with ultra-high sensitivity, high stability, and strong anti-interference capability. It is based on the principle of the limulus amebocyte lysate (LAL) enzymatic reaction and quantitatively detects bacterial endotoxin (also known as lipopolysaccharide or LPS) using colorimetric methods. The detection sensitivity can be as low as 0.01EU/ml, and it can test minute samples, with the sample volume being only 10 microliters or even smaller.
Once this kit is first activated, if stored properly, it remains effective for at least two months. Kits based on LAL for endotoxin testing typically need to be used immediately after preparation, leading to wastage of the reagents. This kit has been carefully optimized to ensure that, once prepared into a solution and stored as recommended in the instructions, it remains effective for at least two months.
This kit offers short detection times and requires a small sample volume. After adding the testing reagent to the sample, the color reaction can be completed in as fast as 16 minutes, enabling easy, quick, and highly sensitive detection of endotoxin content. It doesn't rely on the formation of a clotting protein gel, has strong anti-interference ability, and is suitable for samples with a pale yellow background color, such as plasma and serum. The kit consumes a minimal amount of the sample to be tested, requiring only 10μl per reaction system. It contains azo reagents, ensuring high sensitivity, a broad linear range for sample measurement, and the prevention of interference from the intrinsic color of colored samples.
Usage
Detection of endotoxin contamination in solution samples; Quantitative detection of endotoxin content in various types of samples.
Principle
The limulus reagent is derived from the freeze-dried blood cell lysate of the horseshoe crab species, Limulus polyphemus (Lyophilized Amebocyte Lysate, LAL), or its in vitro recombinant counterpart. This reagent contains factors such as Factor C, Factor B, proclotting enzyme, and clotting protein precursors. Under suitable conditions (temperature, pH, and absence of interfering substances), endotoxins activate Factor C, triggering a series of enzymatic reactions. This leads to the activation of the proclotting enzyme to form the clotting enzyme, which then breaks down the colorant provided in this kit, resulting in the formation of peptides and a yellow compound known as para-nitroaniline (pNA, λmax = 405nm). Over a given period, the amount of pNA generated is directly proportional to the concentration of endotoxins. Concurrently, para-nitroaniline (pNA) can also react with azo reagents to produce a rose-red compound (λmax = 545nm). Based on this, the concentration of endotoxins in the sample to be tested can be quantitatively determined, and interference from the inherent color of the sample at the 405nm absorption peak can be avoided.
Sensitivity
The kit has high sensitivity. The lowest detectable limit can reach 0.01EU. The selectable effective detection ranges are from 0.1E to 1.0EU/ml or 0.01 to 0.10EU/ml.
Storage
Store at 4ºC, valid for two years. After dissolving the endotoxin testing reagent, it should be promptly aliquoted and stored at -20ºC, where it remains valid for one week. If quickly frozen with liquid nitrogen, it can be stored at -80ºC and will be effective for at least two months. After dissolving the remaining reagents, store them at -20ºC, where they remain valid for at least six months.
Precautions
1. The endotoxin testing reagent becomes very unstable once dissolved. It should be used within 10 minutes or stored in aliquots immediately after preparation. Thus, during the use of the kit, all preparations should be made in advance, and the endotoxin testing reagent should only be dissolved or thawed from previously aliquoted frozen stocks just before it is needed.
2. Vessels used in the experiment should be pretreated to remove any potential exogenous endotoxins. A common method is dry baking at 250ºC for at least 60 minutes. Other suitable methods that ensure no interference with bacterial endotoxin testing can also be used. If plasticware is used, select those labeled as endotoxin-free and non-interfering with the experiment.
3. Measures should be taken to prevent microbial contamination during the experimental procedure.
4. If the dissolved and diluted endotoxin standard solution has been left standing for more than 10 minutes, it should be vortexed for 1 minute before use. The thawed endotoxin standard solution should be fully dissolved and vigorously vortexed before being used in subsequent experiments.
5. After dissolving the endotoxin testing reagent from this kit, it should be aliquoted immediately. Store at -20ºC for up to one week or, after quick-freezing with liquid nitrogen, at -80ºC for at least two months. Repeated freeze-thaw cycles must be avoided.
6. For dilution and preparation of samples, we recommend using our endotoxin-free ultrapure water.
7. This product is exclusively for scientific research by professionals. It should not be used for clinical diagnosis or treatment, nor should it be used in food or drugs. It should not be stored in general household areas.
8. For your safety and health, please wear a lab coat and use disposable gloves during the operation.
Only for research and not intended for treatment of humans or animals
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