Multiplex Assay Kit for Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay)

HLF2; LF; GIG12; Lactotransferrin; Growth-inhibiting protein 12; Talalactoferrin; Lfcin-H; Kaliocin-1

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-98 86
EDTA plasma(n=5) 80-97 86
heparin plasma(n=5) 92-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 83-93% 88-95% 80-101% 90-98%
EDTA plasma(n=5) 79-94% 80-102% 88-95% 79-101%
heparin plasma(n=5) 79-91% 79-98% 80-97% 78-90%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:LTF) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
The International Journal of Biochemistry & Cell Biology Changes of hepatic lactoferrin gene expression in two mouse models of the acute phase reaction ScienceDirect: S1357272511002524
BMC Veterinary Research Changes in composition of colostrum of Egyptian buffaloes and Holstein cows BioMed: 17466148
Plos One p38MAPK, ERK and PI3K Signaling Pathways Are Involved in C5a-Primed Neutrophils for ANCA-Mediated Activation PlosOne: source
The Journal of Biological Chemistry Sphingolipid Pathway Regulates Innate Immune Responses at the Fetomaternal Interface during Pregnancy Pubmed:25505239
PLoS One Control of Methicillin-Resistant Staphylococcus aureus Pneumonia Utilizing TLR2 Agonist Pam3CSK4 Pubmed:26974438
Bioscience, Biotechnology, and Biochemistry Salivary lactoferrin is transferred into the brain via the sublingual route pubmed:28351211
Pulmonary Pharmacology & Therapeutics Effects of fluticasone propionate and budesonide on the expression of immune defense genes in bronchial epithelial cells Pubmed:29627483
Microbial Pathogenesis TLR2 agonist Pam3CSK4 enhances the antibacterial functions of GM-CSF induced neutrophils to methicillin-resistant Staphylococcus aureus
Assessment of the Diagnostic Value of IL-10 and Lactoferrin by BIRADS Categories in Breast Neoplasms
J Clin Med A Three-Protein Panel to Support the Diagnosis of Sepsis in Children Pubmed:35329889
Food Funct Three Urban of China: A Cohort Study of Maternal-Infant Factors and HM Protein Components Pubmed:35438093
Biomedicines Myeloid-Specific Pyruvate-Kinase-Type-M2-Deficient Mice Are Resistant to Acute Lung Injury Pubmed:35625931
Catalog No. Related products for research use of Sus scrofa; Porcine (Pig) Organism species Applications (RESEARCH USE ONLY!)
RPA780Po01 Recombinant Lactoferrin (LTF) Positive Control; Immunogen; SDS-PAGE; WB.
APA780Po01 Active Lactoferrin (LTF) Cell culture; Activity Assays.
PAA780Po01 Polyclonal Antibody to Lactoferrin (LTF) WB; IHC; ICC; IP.
LAA780Po81 FITC-Linked Polyclonal Antibody to Lactoferrin (LTF) WB; IHC; ICC; IF.
LAA780Po71 Biotin-Linked Polyclonal Antibody to Lactoferrin (LTF) WB; IHC; ICC.
MAA780Po21 Monoclonal Antibody to Lactoferrin (LTF) WB; IHC; ICC; IP.
SEA780Po ELISA Kit for Lactoferrin (LTF) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA780Po Multiplex Assay Kit for Lactoferrin (LTF) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.