Thermo Fisher Scientific Pierce Rapid Gel Clot User guide

Pierce™ Rapid Gel Clot Endotoxin Assay Kits

Catalog NumbersA43879, A43880, A43881, and A43882

Pub.No. MAN0028592 Rev. A.0

WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and

gloves. Safety Data Sheets (SDSs) are available from thermoﬁsher.com/support.

Product description

The Thermo Scientiﬁc™ Pierce™ Rapid Gel Clot Endotoxin Assay Kits are qualitative tests that use amebocyte lysates derived from the Limulus

horseshoe crab hemolymph to detect endotoxin levels in test samples. The limulus amebocyte lysate (LAL) is widely used as a simple and

sensitive assay for detecting endotoxin lipopolysaccharide found as part of the membrane in gram-negative bacteria. When endotoxin encounters

the amebocyte lysate, a series of enzymatic reactions form a gel-like clot. A clot will form in the sample tube if the amount of endotoxin in the

sample is greater than or equal to the listed sensitivity (in EU/mL) of the kit.

Note: The Pierce™ Rapid Gel Clot Endotoxin Assay Kits are not the standard gel clot method speciﬁed in the USP Chapter 85 Bacterial Endotoxin

Test. This assay kit is not licensed by the FDA and is not to be used for the end-product release of pharmaceutical drugs, devices, or biologics

that are FDA regulated.

Contents and storage

Item Catalog No. Amount Storage

Pierce™ Rapid Gel Clot Endotoxin Assay Kit,

0.03EU/mL A43879 Each kit sucient for 10 tests

Includes:

• 10 each – Sample Reaction Tubes (contains LAL)

• 10 each – Positive Control Tubes (contains endotoxin)

• 10 packs – 1 mL Endotoxin-free Pipette Tips

• 10 each – 10 mL Endotoxin-free Sample Bottles

• Store Sample and Positive

Control Reaction Tubes at

4°C.

• Store pipette tips and

sample bottles at room

temperature.

Pierce™ Rapid Gel Clot Endotoxin Assay Kit,

0.125EU/mL A43880

Pierce™ Rapid Gel Clot Endotoxin Assay Kit,

0.25EU/mL A43881

Pierce™ Rapid Gel Clot Endotoxin Assay Kit,

0.5 EU/mL A43882

Required materials not supplied

• Tube rack

• Non-circulating water bath or heating block incubator set to 37°C ± 1°C

Procedural guidelines

• All materials needed for optimal use of the kit must be endotoxin-free. This includes pipette tips and any containers used to hold the sample to

be tested. Use the 10-mL endotoxin-free glass sample vials for holding samples before gel clot testing.

• Gently add samples to the positive control vials to prevent the formation of foams and bubbles. Gentle mixing and handling are essential

during the entire procedure. Disturbing the vials during the incubation period and test measurement can break the clot forcing an inconclusive

result.

• A heating block set to 37°C can be used, but for best results, a tube rack placed in a non-circulating water bath at 37°C yields more

consistent results with less clot disturbance.

• Each kit lot has a deﬁned assay incubation time as listed in the Certiﬁcate of Analysis. Use the time listed on the Certiﬁcate of Analysis for

step6 in the Assay Procedure below.

Assay procedure

1. Preheat the incubator to 37°C ± 1°C. We recommend measuring the incubator temperature using a calibrated thermometer to ensure that the

incubator is accurate.

2. Collect the test sample in the supplied endotoxin-free sample bottle.

3. Open the Sample and Positive Control tubes. If the aluminum crimp does not separate from the tube, use forceps to remove the crimp.

4. Transfer 0.5 mL of the test sample into the Sample tube using an endotoxin-free pipette tip. Gently mix the tube contents by lightly tapping

the bottom several times to dissolve the LAL reagent.

5. Transfer 0.25 mL of the liquid from the Sample tube to the Positive Control tube. Gently mix the tube contents by lightly tapping the bottom

several times.

6. Immediately after mixing, gently place the Sample and Positive Control tubes in the 37°C water bath. Incubate for the amount of time listed in

the Certiﬁcate of Analysis.

7. Immediately after incubating for the speciﬁc time, gently remove the tubes one at a time from the incubator. Gently invert the tube

180 degrees to read the test results. Perform this action in one smooth motion.

USER GUIDE

For Research Use Only. Not for use in diagnostic procedures.

Interpret assay results

Positive Control Sample Result

+ + The endotoxin concentration of the samples is equal to or higher than the labeled sensitivity of the kit.

+ – The endotoxin concentration of the sample is lower than the labeled sensitivity of the kit.

– + Experiment is invalid. Repeat assay.

– –

Troubleshooting

Observation Possible cause Recommended action

Positive control tube is negative Tube was disturbed during the assay. Repeat the assay with gentle handling of the tubes.

Inhibitory substance was in the

sample.

Optimal sample pH is between 6.0–8.0. If the sample is outside of this

range, add a small amount of 50mM Pierce™ Endotoxin-Free Tris Buer

to bring the sample to the proper pH. Include the dilution caused by the

addition of the Tris buer when determining the endotoxin level of the

sample.

Incubator was not at the proper

temperature.

Use a thermometer to ensure the water bath temperature has equilibrated.

A positive result is not expected Endotoxin was introduced to the

sample during processing.

Maintain aseptic technique and use endotoxin-free plastics and glassware.

1,3-beta-D-glucans were present in

the sample.

Addition of Pierce™ B-glucan Blocker (Cat. No. A43887) to the sample can

help reduce the eect of the 1,3-beta-D-glucans.

Thermo Fisher Scientific Pierce Rapid Gel Clot User guide

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