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Multiplex Assay Kit for Semaphorin 4D (SEMA4D) ,etc. by CBA (Cytometric Bead Array)

CD100; A8; BB18; GR3; M-Sema G; M-Sema-G; SEMAJ; Coll-4; Sema Domain,Immunoglobulin Domain(Ig),Transmembrane Domain(TM)and Short Cytoplasmic Domain 4D

Specificity

This assay has high sensitivity and excellent specificity for detection of Semaphorin 4D (SEMA4D) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Semaphorin 4D (SEMA4D) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Semaphorin 4D (SEMA4D) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Semaphorin 4D (SEMA4D) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-103 93
EDTA plasma(n=5) 84-102 94
heparin plasma(n=5) 80-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Semaphorin 4D (SEMA4D) ,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 Semaphorin 4D (SEMA4D) ,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 Semaphorin 4D (SEMA4D) ,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) 88-99% 92-105% 86-94% 78-101%
EDTA plasma(n=5) 79-97% 80-89% 80-102% 93-104%
heparin plasma(n=5) 80-89% 80-104% 80-104% 79-94%

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#:SEMA4D) 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. 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
Expert Opin Ther Targets. Circulating semaphorin-4D and plexin-B1 levels in postmenopausal women with low bone mass: the 3-month effect of zoledronic acid, denosumab or teriparatide treatment Pubmed:25395071
Obesity (Silver Spring). Genome-wide analysis identifies an african-specific variant in SEMA4D associated with body mass index. pubmed:28296344
Clinical Lymphoma, Myeloma & Leukemia A Revised Staging System for Waldenström's Macroglobulinemia 10.1016/j.clml.2017.07.146
Clinical Lymphoma, Myeloma & Leukemia Extramedullary Multiple Myeloma: Clinical Features and Prognosis in Argentina 10.1016/j.clml.2017.07.144
Blood Cancer Journal Semaphorin 4D correlates with increased bone resorption, hypercalcemia, and disease stage in newly diagnosed patients with multiple myeloma Pubmed:29748532
Analytical Biochemistry A high-sensitivity enzyme immunoassay for the quantification of soluble human semaphorin 4D in plasma Pubmed: 30879960
Journal of Allergy and Clinical Immunology-In Practice Pathological and therapeutic implications of eosinophil-derived semaphorin 4D in eosinophilic chronic rhinosinusitis Pubmed: 32035658
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPB430Hu01 Recombinant Semaphorin 4D (SEMA4D) Positive Control; Immunogen; SDS-PAGE; WB.
PAB430Hu01 Polyclonal Antibody to Semaphorin 4D (SEMA4D) WB
MAB430Hu22 Monoclonal Antibody to Semaphorin 4D (SEMA4D) WB; IHC; ICC; IP.
SEB430Hu ELISA Kit for Semaphorin 4D (SEMA4D) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB430Hu Multiplex Assay Kit for Semaphorin 4D (SEMA4D) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.
CMB430Hu Multiplex Assay Kit for Semaphorin 4D (SEMA4D) ,etc. by CBA (Cytometric Bead Array) FLIA Kit for Antigen Detection.