Multiplex Assay Kit for Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,etc. by FLIA (Flow Luminescence Immunoassay)

CCL3; MIP1-A; SCYA3; Chemokine C-C-Motif Ligand 3; Small Inducible Cytokine A3; Homologous To Mouse Mip-1a; Tonsillar Lymphocyte LD78 Alpha Protein

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-94 89
EDTA plasma(n=5) 93-101 96
heparin plasma(n=5) 89-101 96
sodium citrate plasma(n=5) 78-98 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,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 Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,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 Macrophage Inflammatory Protein 1 Alpha (MIP1a) ,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) 80-96% 98-105% 86-104% 93-102%
EDTA plasma(n=5) 93-101% 91-105% 91-99% 99-105%
heparin plasma(n=5) 88-103% 89-102% 80-103% 91-99%
sodium citrate plasma(n=5) 83-91% 83-90% 92-101% 98-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MIP1a) 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
Inhalation Toxicology Age influence on hypersensitivity pneumonitis induced in mice by exposure to Pantoea agglomerans Pubmed: 24044680
Scientific Reports The Transcriptional Foundations of Sp110-mediated Macrophage (RAW264. 7) Resistance to Mycobacterium tuberculosis H37Ra Pubmed:26912204
ActaMedicaMediterranea CIRCULATING MACROPHAGE INFLAMMATORY PROTEIN-1 ALPHA (MIP1-Α) AS A POTENTIAL BIOMARKER FOR THE DIAGNOSIS OF PATIENTS WITH … 57e97ac908aeb34bc08fd41a.pdf
National Natural Scientific Role of Gut-Derived Endotoxin on Type I Collagen Production in the Rat Pancreas after Chronic Alcohol Exposure pubmed:29121396
Pharmacological Reports Changes in the concentrations of inflammatory and oxidative status biomediators (MIP-1 α, PMN elastase, MDA, and IL-12) in depressed patients with and without posttraumatic stress disorder 10.1016/j.pharep.2017.08.008
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Journal of Immunology Research Therapeutic Potential of Pien Tze Huang on Experimental Autoimmune Encephalomyelitis Rat Pubmed:29682587
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Pharmacological Reports Changes in the concentrations of inflammatory and oxidative status biomediators (MIP-1 α, PMN elastase, MDA, and IL-12) in depressed patients with and without … Pubmed:29339257
Journal of Food Science CQPC06 Activity Prevents Dextran Sulfate Sodium‐Induced Colitis by Regulating the IL‐8 Pathway Doi: 10.1111/1750-3841.14346
Biochemical and Biophysical Research Communications The anti-inflammatory and anti-oxidative effects of conbercept in treatment of macular edema secondary to retinal vein occlusion Pubmed: 30558792
LIFE SCIENCES Intravitreal conbercept improves outcome of proliferative diabetic retinopathy through inhibiting inflammation and oxidative stress Pubmed: 33227274
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Life Sci PIGU promotes hepatocellular carcinoma progression through activating NF-κB pathway and increasing immune escape Pubmed: 32971102
Nature Communications A vaccine-based nanosystem for initiating innate immunity and improving tumor immunotherapy Pubmed: 32332752
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