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Lal assay

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The LAL (Limulus Amebocyte Lysate) assay is a laboratory test used to detect and quantify the presence of endotoxins, a component of the cell wall of Gram-negative bacteria. The assay utilizes the clotting reaction that occurs when endotoxins interact with the lysate of amebocytes, a type of blood cell found in the horseshoe crab. The resulting clot formation is measured and correlated to the endotoxin concentration in the sample.

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Lab products found in correlation

Sandwich elisa kit, by BioLegend (1 mentions) Reg3α, by R&D Systems (1 mentions) Soluble cd163, by R&D Systems (1 mentions) Scd14, by R&D Systems (1 mentions) I fabp, by R&D Systems (1 mentions)

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Pyrochrome LAL assay (4 citations) Fungitell ® LAL assay (3 citations) Chromo-LAL assay (2 citations)

6 protocols using lal assay

1

Mouse Experiments with TLR9 Agonist and Inhibitor

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The single-stranded ODNs of the TLR9 agonist CpG ODN (CpG 5805), TLR9 inhibitor CCT ODN and TGF-β2 ASO TIO3 were used in mouse experiments.41 (link),62 (link) All ODNs were synthesized, purified and fully phosphorothioate-modified by Takara Biotech (Dalian, China). The ODNs were diluted in PBS and no detectable endotoxin was detected by the LAL assay conducted by Associates of Cape Cod, Inc. (Massachusetts, USA).
Zhu M., Wang S., Qu K., Lu F., Kou M., Yao Y., Zhu T., Yu Y., Wang L, & Yan C. (2024). The trogocytosis of neutrophils on initial transplanted tumor in mice. iScience, 27(5), 109661.
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2

Purification of A-rRBD and B-rRBD Proteins

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The purification of A-rRBD and B-rRBD expressed in E. coli JM109 (DE3) strain have been previously described [30 , 31 (link)]. All purification steps were analyzed by 8 % SDS-PAGE. The residual endotoxin was determined using the Limulus amoebocyte lysate (LAL) assay (Associates of Cape Cod, Inc., Cape Cod, MA).
Huang J.H., Wu C.W., Lien S.P., Leng C.H., Hsiao K.N., Liu S.J., Chen H.W., Siu L.K, & Chong P. (2015). Recombinant lipoprotein-based vaccine candidates against C. difficile infections. Journal of Biomedical Science, 22(1), 65.
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3

Quantifying Plasma Endotoxin Levels

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Plasma endotoxin contents were determined using the Diazo-coupled limulus amebocyte lysate (LAL) assay (Associates of Cape Cod, Inc., E. Falmouth, MA, USA) according to the manufacturer’s protocol. The assay reagents-treated samples lead to the formation of a magenta derivative that absorbs light at 545 nm.
Song J.H., Kim H., Jeong M., Kong M.J., Choi H.K., Jun W., Kim Y, & Choi K.C. (2021). In Vivo Evaluation of Dendropanax morbifera Leaf Extract for Anti-Obesity and Cholesterol-Lowering Activity in Mice. Nutrients, 13(5), 1424.
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4

Recombinant Reference Allergen Production

Cited 2 times
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Recombinant reference allergens (Bet v 1.0101, Bet v 1.0102, Aln g 1.0101, Cor a 1.0104, Car b 1.0109 and Que a 1.0301) were produced according to published methods [9 (link), 10 (link)] and characterized physico-chemically [6 (link)]. Endotoxin content was < 0.3 ng/mg for Bet v 1.0101, < 0.3 ng/mL for Bet v 1.0102, < 0.3 ng/mL for Aln g 1.0101, 0.8 ng/mL for Cor a 1.0104, < 0.3 ng/mg for Car b 1.0109, and < 0.3 ng/mg for Que a 1.0301 as determined by limulus amoebocyte lysate (LAL) assay (Associates of Cape Cod, Inc., East Falmouth, MA, USA).
Pichler U., Hauser M., Hofer H., Himly M., Hoflehner E., Steiner M., Mutschlechner S., Hufnagl K., Ebner C., Mari A., Briza P., Bohle B., Wiedermann U., Ferreira F, & Wallner M. (2014). Allergen hybrids – next generation vaccines for Fagales pollen immunotherapy. Clinical and Experimental Allergy, 44(3), 438-449.
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5

Cytokine Quantification in oxLDL-Stimulated DCs

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The amounts of IL-17, IFNγ, or IL-6 in cultured supernatant or serum were measured by using sandwich ELISA kit (BioLegend). In some experiments, primary DCs were freshly isolated from the spleens of C57BL/6, Myd88−/−, Tlr4−/−, Cd36−/− mice by using anti-CD11c-beads. The isolated primary DCs and BMDCs (1×106 cells/ml) were stimulated with 100 ng/ml LPS, 20 µg/ml LDL, or three types of oxLDLs with differential oxidation extent (low Tbars (5–15), medium Tbars (25–35), high Tbars (50–70); all from Biomedical Technologies). The endotoxin levels of oxLDLs and LDL were <0.5 EU/mg LDL, as measured by an LAL assay (Associates of Cape Cod). Twenty-four hours after stimulation, the levels of IL-6 in the supernatant were measured by ELISA. The oxLDL levels in mouse serum were measured by using ELISA kit (Cusabio Biotech) according to the manufacturer’s instruction.
Lim H., Kim Y.U., Sun H., Lee J.H., Reynolds J.M., Hanabuchi S., Wu H., Teng B.B, & Chung Y. (2014). Proatherogenic conditions promote autoimmune T helper 17 cell responses in vivo. Immunity, 40(1), 153-165.
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6

Plasma biomarkers of gut barrier dysfunction

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Sequential blood samples from a total of 120 participants were analyzed. Plasma βDG was measured by the Fungitell Limulus Amebocyte Lysate (LAL) assay (Associates of Cape Cod, Inc, East Falmouth, MA, USA) according to the manufacturer's instruction. Enzyme-linked immunosorbent assays (ELISAs) were performed to quantify plasma LPS (CUSABIO, Wuhan, Hubei, China), LPS binding protein (LBP, Hycultbiotech, Uden, Netherlands), soluble LPS receptor CD14 (sCD14), intestinal fatty acid binding protein (IFABP), regenerating islet-derived protein 3α (REG3α), and soluble CD163 (R&D Systems, Minneapolis, MN, USA).
Xun J., Guo S., Xu Y., Chen R., Tang Q., Zhang X., Liu D., Zhang R., Shen Y., Liu L., Wan J., Chen J, & Lu H. (2022). Circulating (1 → 3)-β-D-Glucan as an immune activation marker decreased after ART in people living with HIV. Frontiers in Public Health, 10, 981339.
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