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Human ll 37 elisa kit

Manufactured by Hycult Biotech
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Sourced in Germany, United States

The Human LL-37 ELISA kit is a quantitative enzyme-linked immunosorbent assay (ELISA) designed to measure the concentration of the human cathelicidin antimicrobial peptide LL-37 in various biological samples. The kit utilizes a specific antibody to capture and detect the target analyte.

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Lab products found in correlation

Hnp1 3 human elisa kit, by Hycult Biotech (1 mentions) Creatinine microplate assay, by Oxford Biomedical Research (1 mentions) Abts elisa development kit, by Thermo Fisher Scientific (1 mentions) Human myeloperoxidase duoset elisa kit, by R&D Systems (1 mentions) Cobas c501 analyzer, by Roche (1 mentions) Recombinant ll 37, by AnaSpec (1 mentions) Agar, by Thermo Fisher Scientific (1 mentions) Tryptic soy broth (tsb), by Acumedia (1 mentions) Fibroblast growth factor 2 fgf 2, by R&D Systems (1 mentions) Cli 095, by InvivoGen (1 mentions) Oxpapc, by InvivoGen (1 mentions) 1 25 dihydroxyvitamin d3, by Cayman Chemical (1 mentions) Ll 37 peptide, by AnaSpec (1 mentions) Monophosphoryl lipid a mpla, by InvivoGen (1 mentions) Gw0742, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Trypsin 2 s, by Merck Group (1 mentions) Ampicillin amp, by Merck Group (1 mentions) Collagenase type 1, by Worthington (1 mentions) Liaison 25 oh vitamin d total, by DiaSorin (1 mentions)

14 protocols using human ll 37 elisa kit

1

Biomarker Detection in Urine and Serum

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Study staff identified patients in real time on weekdays from 8AM to 9PM. Patients completed an initial survey covering demographics, symptoms, and medical history, and provided urine and blood samples. Electronic medical record chart review (EPIC, EPIC Systems Corp., Verona, WI) was used to record vital signs, diagnostic study results, and disposition.
Urine was collected using standard clinical procedures (clean-catch or catheter). Both urine and serum were centrifuged and stored at −80°C. Enzyme-linked immunosorbent assays (ELISA) were performed using HNP1-3 Human ELISA Kit (Hycult Biotech-Plymouth Meeting, PA), Defensin-5 ELISA kit (Lifeome Biolabs-Oceanside, CA), Human Beta Defensin 2 ELISA Kit (Lifeome Biolabs), and LL-37 Human Elisa Kit (Hycult Biotech). For all but HNP 1-3, ELISA results were divided by urine creatinine (measured using the Oxford Biomedical Research creatinine microplate assay, Rochester Hills, MI) to standardize for urine concentration. Samples were run in duplicate. Serum 25-hydroxyvitamin D (25OHD) levels were measured by liquid chromatography-tandem mass spectrometry.
Caterino J.M., Hains D.S., Camargo CA J.r., Quraishi S.A., Saxena V, & Schwaderer A.L. (2015). A prospective, observational pilot study of the use of urinary antimicrobial peptides in diagnosing emergency department patients with positive urine cultures. Academic emergency medicine : official journal of the Society for Academic Emergency Medicine, 22(10), 1226-1230.
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2

LL37 Quantification Using ELISA

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LL37 levels were analyzed using the LL37 Human ELISA kit (HK321, Hycult Biotech, Uden, The Netherlands) according to the manufacturer’s instructions. Briefly, thawed samples were centrifuged, and the supernatant was transferred to antibody-coated microtiter wells to capture LL37, incubated with a biotinylated secondary antibody and detected by streptavidin-peroxidase.
Gyll J., Ridell K., Öhlund I., Karlsland Åkeson P., Johansson I, & Lif Holgerson P. (2018). Vitamin D status and dental caries in healthy Swedish children. Nutrition Journal, 17, 11.
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3

Quantification of HBD1 and LL-37 Peptides

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HBD1 and LL-37 peptide release was quantified in supernatants from 25-day long-term cultured A549 cells using a human BD-1 standard ABTS ELISA development kit (PeproTech, Hamburg, Germany) and LL-37 human ELISA kit (HycultBiotech, Beutelsbach, Germany) according to the manufacturers' instructions.
Tran H.T., Peterburs P., Seibel J., Abramov-Sommariva D, & Lamy E. (2022). In vitro Screening of Herbal Medicinal Products for Their Supportive Curing Potential in the Context of SARS-CoV-2. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 8038195.
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4

Comprehensive Biomarker Panel Analysis

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Complete blood count with differential leukocyte count was assessed using a Sysmex XN-1000™ Hematology Analyzer (Sysmex Group’s, Kobe, Japan). Serum creatinine, BUN, uric acid, total proteins, albumin, bilirubin, C-reactive protein, natrium, potassium, calcium and phosphorus were analyzed using standard laboratory methods (Cobas c501 analyzer, Roche Diagnostic, Basel, Switzerland).
Urinary creatinine was measured using Jaffé reaction as described previously (21 (link)). Briefly, 10 microliters of samples were mixed with 200 microliters of freshly prepared reagent consisted of 0.2 M sodium hydroxide and 25 mM picric acid (5:1 ratio). Absorbance was measured at 492 nm. Absorbance measured after 1st minute was subtracted from the absorbance measured at 6th minute.
Concentration of myeloperoxidase (MPO) in urine was determined using commercially available spectrophotometric assay according to manufactureŕs protocol (Human Myeloperoxidase DuoSet Elisa Kit, R&D Systems, Minneapolis, MN, United States). Absorbance was measured at 450 nm. Limit of detection was 62.5 pg/ml.
Similarly, cathelicidin in urine was measured by commercial Elisa kit as recommended by the manufactureŕs protocol (LL-37 Human Elisa kit, Hycult Biotech, Wayne, PA, United States). Urine samples were 10-times diluted. Absorbance was measure at 450 nm. Limit of detection was 0.14 ng/ml.
Krivošíková K., Šupčíková N., Gaál Kovalčíková A., Janko J., Pastorek M., Celec P., Podracká Ľ, & Tóthová Ľ. (2023). Neutrophil extracellular traps in urinary tract infection. Frontiers in Pediatrics, 11, 1154139.
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5

Quantification of HBD1 and LL-37 in A549 Cells

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HBD1 and LL-37 peptide release was quantified in supernatants from 25 day long-term cultured A549 cells using a human BD-1 Standard ABTS ELISA Development Kit (PeproTech, Germany) and LL-37 human ELISA kit (HycultBiotech, Germany) according to the manufacturer's instructions.
Tran H.T.T., Peterburs P., Seibel J., Abramov-Sommariva D., & Lamy E. (2021). In vitroscreening of herbal medicinal products for their supportive curing potential in the context of SARS-CoV-2.
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6

Evaluating LL37-mediated EV Immunomodulation

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EVs from active BD, inactive BD and healthy subjects were incubated with healthy PBMCs and autologous PBMCs (1.25 × 106 per ml) for 12 h at a ratio of 1:2 (Cell:EV) in 96-well flat bottom culture plates. In some experiments, healthy EVs were mixed with recombinant LL37 (AnaSpec, Freemont, CA, USA) overnight and incubated with autologous PBMCs ifor 12h. In neutralization studies anti-LL37 mAb (HyCult Biotech, Uden, Netherlands) were incubated with EV treated PBMCs at indicated concentrations. LPS (5 µg ml–1) and PGN (5 µg ml–1) were used as positive controls throughout these assays. Culture supernatants were collected and IL6 production was determined by ELISA.[34 (link)] The concentration of LL37 from plasma and EV samples was measured by a human LL37 ELISA kit (HyCult Biotech), according to the manufacturer’s protocol.
Kahraman T., Gucluler G., Simsek I., Yagci F.C., Yildirim M., Ozen C., Dinc A., Gursel M., Ikromzoda L., Sutlu T., Gay S, & Gursel I. (2017). Circulating LL37 targets plasma extracellular vesicles to immune cells and intensifies Behçet’s disease severity. Journal of Extracellular Vesicles, 6(1), 1284449.
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7

Quantifying LL-37, S1P, and LCN2 in Cells

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A human LL-37 ELISA kit (Hycult Biotech, Wayne, PA, USA and a human S1P ELISA kit (MyBioSource, San Diego, CA, USA) were used to determine LL-37 (the human cathelicidin peptide) and S1P levels in hMCs or NHEKs in accordance with the manufacturer’s instructions. Mouse and Human LCN2 ELISA kits (R&D Systems) were used for quantifying LCN2 protein secretion in both hMCs and mMCs.
Chang Y.L., Wang Z., Igawa S., Choi J.E., Werbel T, & Di Nardo A. (2019). Lipocalin 2: A New Antimicrobial in Mast Cells. International Journal of Molecular Sciences, 20(10), 2380.
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8

Isolation and Culture of Human Immune Cells

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Dulbecco’s modified Eagle’s medium (DMEM), α-Minimum Essential Medium (α-MEM), antibiotic-antimycotic (anti-anti), fetal bovine serum (FBS), and phosphate-buffered saline (PBS) were purchased from the Life Technologies (Grand Island, NY). OxPAPC, CLI-095, and monophosphoryl lipid A (MPLA) were purchased from the Invivogen (San Diego, CA). Fibroblast growth factor-2 (FGF-2) was obtained from the R&D System (Minneapolis, MN). Tryptic soy broth was purchased from the Acumedia (Lansing, MI). Agar was obtained from the Fisher Scientific (Pittsburgh, PA). Human LL-37 ELISA kit was purchased from the Hycult Biotech (Plymouth Meeting, PA). LL-37 peptide was purchased from the AnaSpec (Fremont, CA). GW0742, trypsin IIS, and ampicillin (Amp) were purchased from the Sigma-Aldrich (St. Louis, MO). Collagenase type I was purchased from the Worthington (Lakewood, NJ). 1,25-dihydroxy vitamin D3 was purchased from the Cayman Chemical (Ann Arbor, MI).
Yagi H., Chen A.F., Hirsch D., Rothenberg A.C., Tan J., Alexander P.G, & Tuan R.S. (2020). Antimicrobial activity of mesenchymal stem cells against Staphylococcus aureus. Stem Cell Research & Therapy, 11, 293.
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9

Lipigenine Increases Cathelicidin (LL-37) Levels

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EXAMPLE 7

A topical composition with Lipigenine™ was tested for its ability to increase concentration of Cathelicidin (LL37), an amphipathic alpha-helical peptide that plays an important role in defense against local infection and invasion of pathogens at sites of inflammation and wounds. The human LL-37 ELISA kit was obtained from Hycult Biotech (Cat#HK321). ELISA were performed according to the manufactory instructions of each kit by adding 100 μl/well of culture medium after overnight treatment. The results were measured using a colorimeter, absorbance was measured at 450 nanometers (nm) within 30 minutes. Wavelength correction was set to 570 nm.

The addition of Lipigenine™ showed increased LL-37 concentration at both 0.1% and 1% Lipigenine™ in solution as compared to the control. An increase in LL-37 concentration of 32% for a 0.1% Lipigenine™ formulation while an increase in LL-37 concentration of 38% was observed for a 1% Lipigenine™ formulation. These results are shown in FIG. 7.

US10806769B2. Antimicrobial peptide stimulating cleansing composition (2020-10-20). GOJO Industries, Inc. [US]. Inventors: Kegui Tian [US], Jessica Rae Tittl [US], Venkatesan V. Padyachi [US].
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10

Lipigenine Increases Antimicrobial Peptide LL-37

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Example 7

A topical composition with Lipigenine™ was tested for its ability to increase concentration of Cathelicidin (LL37), an amphipathic alpha-helical peptide that plays an important role in defense against local infection and invasion of pathogens at sites of inflammation and wounds. The human LL-37 ELISA kit was obtained from Hycult Biotech (Cat #HK321). ELISA were performed according to the manufactory instructions of each kit by adding 100 μl/well of culture medium after overnight treatment. The results were measured using a colorimeter, absorbance was measured at 450 nanometers (nm) within 30 minutes. Wavelength correction was set to 570 nm.

The addition of Lipigenine™ showed increased LL-37 concentration at both 0.1% and 1% Lipigenine™ in solution as compared to the control. An increase in LL-37 concentration of 32% for a 0.1% Lipigenine™ formulation while an increase in LL-37 concentration of 38% was observed for a 1% Lipigenine™ formulation. These results are shown in FIG. 7.

US11633451B2. Antimicrobial peptide stimulating cleansing composition (2023-04-25). GOJO Industries, Inc. [US]. Inventors: Kegui Tian [US], Jessica Rae Tittl [US], Venkatesan V. Padyachi [US].
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