ELISA Kit for Cathelicidin Antimicrobial Peptide (CAMP)

CAP18; FALL39; HSD26; LL37; 18 kDa Cationic Antimicrobial Protein; Antibacterial protein FALL-39; FALL-39 peptide antibiotic

Specificity

This assay has high sensitivity and excellent specificity for detection of Cathelicidin Antimicrobial Peptide (CAMP).
No significant cross-reactivity or interference between Cathelicidin Antimicrobial Peptide (CAMP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Cathelicidin Antimicrobial Peptide (CAMP) and the recovery rates were calculated by comparing the measured value to the expected amount of Cathelicidin Antimicrobial Peptide (CAMP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-103 98
EDTA plasma(n=5) 95-103 99
heparin plasma(n=5) 79-92 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cathelicidin Antimicrobial Peptide (CAMP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cathelicidin Antimicrobial Peptide (CAMP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cathelicidin Antimicrobial Peptide (CAMP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 78-92% 88-95% 96-103% 91-101%
EDTA plasma(n=5) 80-103% 92-105% 89-105% 80-93%
heparin plasma(n=5) 80-102% 87-96% 95-104% 99-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
University of Waikato The Assessment of Vitamin D, Antimicrobial Peptides and Procalcitonin in Bronchiectasis Waikato:Source
Clin Infect Dis. Host immune response to tuberculous meningitis Pubmed:25301213
Vaccine On birth single dose live attenuated OPV and BCG vaccination induces gut cathelicidin LL37 responses at 6 week of age: A natural experiment Pubmed:25444792
Urology Modulating the Internalization of Bacille Calmette-Guérin by Cathelicidin in Bladder Cancer Cells Pubmed:25681250
Urology Modulating the Internalization of Bacille Calmette-Guérin by Cathelicidin in Bladder Cancer Cells PubMed: 25681250
Sarcoidosis Vasc Diffuse Lung Dis Cathelicidin as a link between sarcoidosis and tuberculosis PubMed: 26422567
Allergy and Asthma Proceedings The association of vitamin D, cathelicidin, and vitamin D binding protein with acute asthma attacks in children PubMed: 26108071
J Clin Microbiol. Swiftly Decreasing Cerebrospinal Fluid Cathelicidin Concentration Predicts Improved Outcome in Childhood Bacterial Meningitis. Pubmed:27008883
experimental dermatology Light-emitting diodes downregulate cathelicidin, kallikrein and toll-like receptor 2 expressions in keratinocytes and rosacea-like mouse skin. pubmed:27315464
PLOS ONE Reduced Expression of the Extracellular Calcium-Sensing Receptor (CaSR) Is Associated withActivation of the Renin-Angiotensin System (RAS) to Promote Vascular Remodeling in the Pathogenesis of Essential Hypertension. pubmed:27391973
Front Pharmacol.  Ginsenoside Rg5 Inhibits Succinate-Associated Lipolysis in Adipose Tissue and Prevents Muscle Insulin Resistance. pubmed:28261091
Allergologia et immunopathologia The relation of innate and adaptive immunity with viral-induced acute asthma attacks: Focusing on IP-10 and cathelicidin pubmed:27955890
Chinese Journal of Osteoporosis and Bone Mineral Research Effects of cold exposure on the bone marrow adipose tissue DOI: 10.3969/j.issn.1674-2591.2017.05.008
Scientific Reports Measurements of AMPs in stratum corneum of atopic dermatitis and healthy skin–tape stripping technique Pubmed:29374283
Experimental Parasitology Antimalarial activity of vitamin D3 (VD3) does not result from VD3-induced antimicrobial agents including nitric oxide or cathelicidin Pubmed: 30904694
Theranostics Myeloid cell-derived LL-37 promotes lung cancer growth by activating Wnt/β-catenin signaling Pubmed: 31149039
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Short-Term versus Long-Term Culture of A549 Cells for Evaluating the Effects of Lipopolysaccharide on Oxidative Stress, Surfactant Proteins and Cathelicidin LL-37 Pubmed: 32050475
MICROB PATHOGENESIS Hypovitaminosis D and reduced cathelicidin are strongly correlated during the multidrug therapy against leprosy Pubmed: 32645421
International Journal of Nanomedicine Effect of Lonicerae japonicae Flos Carbonisata-Derived Carbon Dots on Rat Models of Fever and Hypothermia Induced by Lipopolysaccharide Pubmed: 32606669
博士论文 Relações causais entre a expressão dos genes do receptor de Vitamina D e do Peptídeo Antimicrobiano Catelicidina sobre marcadores sorológicos de pessoas com …
PLoS One Beneficial impact of cathelicidin on hypersensitivity pneumonitis treatment¡ªIn vivo studies 33999928
New Med Analysis of the level of non-specific and specific immunity parameters in saliva of children with osteogenesis imperfecta and study of relationships between?¡­
Type I interferons link skin-associated dysbiotic commensal bacteria to pathogenic inflammation and angiogenesis in rosacea
Int J Mol Sci Compartmentalized Innate Immune Response of Human Fetal Membranes against Escherichia coli Choriodecidual Infection Pubmed:35328414
Cell Rep LL-37 transports immunoreactive cGAMP to activate STING signaling and enhance interferon-mediated host antiviral immunity Pubmed:35649354
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