Multiplex Assay Kit for Annexin A1 (ANXA1) ,etc. by FLIA (Flow Luminescence Immunoassay)

ANX-A1; ANX1; LPC1; Lipocortin I; Chromobindin-9; Calpactin II; Phospholipase A2 inhibitory protein

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Annexin A1 (ANXA1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Annexin A1 (ANXA1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Annexin A1 (ANXA1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Annexin A1 (ANXA1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-99 95
EDTA plasma(n=5) 87-102 96
heparin plasma(n=5) 86-104 90
sodium citrate plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Annexin A1 (ANXA1) ,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 Annexin A1 (ANXA1) ,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 Annexin A1 (ANXA1) ,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) 88-104% 78-95% 80-94% 95-102%
EDTA plasma(n=5) 86-101% 79-99% 80-90% 84-104%
heparin plasma(n=5) 78-101% 79-88% 81-104% 85-98%
sodium citrate plasma(n=5) 90-101% 93-102% 84-93% 90-101%

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#:ANXA1) 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
Plos one Dysregulation of Anti-Inflammatory Annexin A1 Expression in Progressive Crohns Disease PubMed: PMC3794972
African Journals Online Modulatory Effect of Phytoestrogens and Curcumin on Induction of Annexin 1 in Human Peripheral Blood Mononuclear Cells and their Inhibitory Effect on Secretory Phospholipase A2 Ajol: Source
Antimicrobial Agents and Chemotherapy Host Biomarkers of Invasive Pulmonary Aspergillosis to Monitor Therapeutic Response Asm: Source
J Proteomics Characterization of secretomes from a human blood brain barrier endothelial cells in-vitro model after ischemia by stable isotope labeling with aminoacids in cell culture (SILAC) PubMed: 26718731
Diabetes Annexin A1 Is a Key Modulator of Mesenchymal Stromal Cell–Mediated Improvements in Islet Function PubMed: 26470781
PLoS One Annexin A1 Is Increased in the Plasma of Preeclamptic Women PubMed: 26398190
中国现代医学杂志 China Journal of Modern Medicine 膜联蛋白A1 和CA19-9 在胆管癌中的表达及其临床意义 article:8686
arteriosclerosis thrombosis and vascular biology Protective Aptitude of Annexin A1 in Arterial Neointima Formation in Atherosclerosis-Prone Mice-Brief Report. pubmed:28062503
Clinical Science Mesenchymal stromal cells improve human islet function through released products and extracellular matrix. pubmed:29101297
Biomedicine & Pharmacotherapy Proresolving protein Annexin A1: The role in type 2 diabetes mellitus and obesity Pubmed:29677533
 International Journal of Chronic Obstructive Pulmonary Disease Annexin A1 is elevated in patients with COPD and affects lung fibroblast function Pubmed:29440885
Medical Mycology Applying host disease status biomarkers to therapeutic response monitoring in invasive aspergillosis patients Pubmed:29370415
DEVELOPMENTAL NEUROPSYCHOLOGY EFFECTS OF MULTICOMPONENT EXERCISE ON PROGRESSION OF BRAIN ATROPHY AND COGNITION IN AMNESTIC MILD COGNITIVE IMPAIRMENT … pdf:3395
Multiple sclerosis and related disorders Resolution of inflammation in neuromyelitis optica spectrum disorders Pubmed: 30300851
Acta Pharmaceutica Effects of annexin A1 on apoptosis and cell cycle arrest in human leukemic cell lines
npj Science of Food Annexin A1 accounts for an anti-inflammatory binding target of sesamin metabolites Pubmed: 32133417
FASEB JOURNAL ANXA1 directs Schwann cells proliferation and migration to accelerate nerve regeneration through the FPR2/AMPK pathway Pubmed: 32908567
Rev Soc Bras Med Trop Expression of annexin-A1 in blood and tissue leukocytes of leprosy patients 33263684
science advances Annexin-A1 SUMOylation regulates microglial polarization after cerebral ischemia by modulating IKK¦Á stability via selective autophagy 33523920
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
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