Multiplex Assay Kit for Laminin Gamma 2 (LAMC2) ,etc. by FLIA (Flow Luminescence Immunoassay)

B2T; EBR2; EBR2A; LAMB2T; LAMNB2; Kalinin/nicein/epiligrin 100 kDa subunit; Cell-scattering factor 140 kDa subunit; Ladsin 140 kDa; Large adhesive scatter factor 140 kDa

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Laminin Gamma 2 (LAMC2) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Laminin Gamma 2 (LAMC2) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Laminin Gamma 2 (LAMC2) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Laminin Gamma 2 (LAMC2) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-91 88
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 78-92 88
sodium citrate plasma(n=5) 95-103 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Laminin Gamma 2 (LAMC2) ,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 Laminin Gamma 2 (LAMC2) ,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 Laminin Gamma 2 (LAMC2) ,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) 87-101% 86-101% 99-105% 78-96%
EDTA plasma(n=5) 81-91% 90-97% 82-94% 81-96%
heparin plasma(n=5) 98-105% 78-98% 96-103% 78-90%
sodium citrate plasma(n=5) 83-101% 93-103% 89-103% 96-103%

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#:LAMC2) 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
Molecular & Cellular Proteomics LAMC2: A promising new pancreatic cancer biomarker identified by proteomic analysis of pancreatic adenocarcinoma tissues Mcponline: 023507
Clinical Cancer Research Validation of Biomarkers That Complement CA19.9 in Detecting Early Pancreatic Cancer Pubmed:25239611
British journal of cancer Serum LAMC2 enhances the prognostic value of a multi-parametric panel in non-small cell lung cancer PubMed: 26180921
U.S. National Library of Medicine Prognostic significance of circulating laminin gamma2 for early-stage non-small-cell lung cancer pubmed:27462170
American Journal of Obstetrics & Gynecology MFM PRETERM BIRTH PREDICTION IN ASYMPTOMATIC WOMEN AT MID-GESTATION USING A PANEL OF NOVEL PROTEIN BIOMARKERS: The Prediction of PreTerm …
Biomarkers of Preterm Birth
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