ELISA Kit for Annexin A1 (ANXA1)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-101 93
EDTA plasma(n=5) 95-103 99
heparin plasma(n=5) 85-105 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Annexin A1 (ANXA1) 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) 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) 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) 79-94% 91-102% 80-89% 88-102%
EDTA plasma(n=5) 78-94% 86-95% 81-95% 80-92%
heparin plasma(n=5) 83-97% 89-104% 95-103% 92-101%

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
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
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
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