Multiplex Assay Kit for Calreticulin (CALR) ,etc. by CBA (Cytometric Bead Array)

CRT; RO; SSA; CC1qR; ERp60; HACBP; grp60; CRTC; CRP55; Calregulin; Sicca Syndrome Antigen A; Autoantigen Ro; Endoplasmic reticulum resident protein 60

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 95-104 101
EDTA plasma(n=5) 83-104 99
heparin plasma(n=5) 82-93 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Calreticulin (CALR) ,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 Calreticulin (CALR) ,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 Calreticulin (CALR) ,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) 87-102% 92-105% 80-103% 98-105%
EDTA plasma(n=5) 84-101% 78-90% 78-89% 95-102%
heparin plasma(n=5) 84-93% 95-105% 99-105% 79-104%

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
Journal of Investigative Dermatology Oxidative Stress-Induced Calreticulin Expression and Translocation: New Insights into the Destruction of Melanocytes Pubmed: 23771121
International Journal of Molecular Sciences The Anticoagulant Effect of PGI2S and tPA in Transgenic Umbilical Vein Endothelial Cells Is Linked to Up-Regulation of PKA and PKC Mdpi: Source
The Journal of Maternal-Fetal & Neonatal Medicine Amniotic fluid calreticulin in pregnancies complicated by the preterm prelabor rupture of membranes Pubmed:26953684
journal of maternal-fetal & neonatal medicine Cervical fluid calreticulin and cathepsin-G in pregnancies complicated by preterm prelabor rupture of membranes. pubmed:28152632
Talanta Investigation of the interaction between calreticulin and immunoglobulin G by capillary electrophoresis and computer modeling Doi: 10.1016/j.talanta.2018.11.098
Sci Adv Thermostable ionizable lipid-like nanoparticle (iLAND) for RNAi treatment of hyperlipidemia Pubmed:35171673
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