Multiplex Assay Kit for Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array)

AMF; NLK; PHI; SA36; Phosphoglucose Isomerase; Autocrine motility factor; Neuroleukin; Phosphohexose isomerase; Sperm antigen 36

Specificity

This assay has high sensitivity and excellent specificity for detection of Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-94 90
EDTA plasma(n=5) 89-97 93
heparin plasma(n=5) 81-96 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array) 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 Glucose 6 Phosphate Isomerase (GPI) ,etc. by CBA (Cytometric Bead Array) 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) 78-101% 87-95% 81-97% 85-104%
EDTA plasma(n=5) 85-94% 83-101% 78-98% 82-104%
heparin plasma(n=5) 84-99% 90-99% 85-98% 96-105%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Oncology Reports Hyperthermia reduces migration of osteosarcoma by suppression of autocrine motility factor PubMed: 23027359
Arthritis & Rheumatology Bruton's tyrosine kinase deficiency inhibits autoimmune arthritis but fails to block immune complex‐mediated inflammatory arthritis Pubmed:26945549
Rheumatoid Arthritis Bruton's Tyrosine Kinase Deficiency Inhibits Autoimmune Arthritis in Mice but Fails to Block Immune Complex–Mediated Inflammatory Arthritis pubmed:26945549
Infection and Immunity The GPI transamidase complex subunit PbGPI16 of Plasmodium berghei is important for inducing experimental cerebral malaria Pubmed:29784863
Journal of Proteomics Proteomics analysis of asthenozoospermia and identification of glucose-6-phosphate isomerase as an important enzyme for sperm motility Pubmed: 31394311
Scientific Reports The Asthma-associated PER1-like domain-containing protein 1 (PERLD1) Haplotype Influences Soluble Glycosylphosphatidylinositol Anchor Protein (sGPI … Pubmed: 31959860
JOURNAL OF IMMUNOLOGY T–B Lymphocyte Interactions Promote Type 1 Diabetes Independently of SLAM-Associated Protein Pubmed: 33199538
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