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ELISA Kit for Semaphorin 7A (SEMA7A)

CD108; CDw108; H-SEMA-K1; H-Sema-L; JMH; SEMAK1; SEMAL; Semaphorin 7A,GPI Membrane Anchor; John Milton Hagen Blood Group; Semaphorin-K1; Semaphorin-L

Specificity

This assay has high sensitivity and excellent specificity for detection of Semaphorin 7A (SEMA7A).
No significant cross-reactivity or interference between Semaphorin 7A (SEMA7A) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Semaphorin 7A (SEMA7A) and the recovery rates were calculated by comparing the measured value to the expected amount of Semaphorin 7A (SEMA7A) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-92 85
EDTA plasma(n=5) 94-105 97
heparin plasma(n=5) 78-94 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Semaphorin 7A (SEMA7A) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Semaphorin 7A (SEMA7A) 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 7A (SEMA7A) 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) 83-94% 95-102% 79-93% 97-105%
EDTA plasma(n=5) 90-102% 78-99% 99-105% 97-105%
heparin plasma(n=5) 91-98% 94-104% 78-89% 85-99%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
PLOS ONE Semaphorin 7A Aggravates Pulmonary Inflammation during Lung Injury pmc:PMC4720127
Experimental and Therapeutic Medicine Anti-Semaphorin-7A single chain antibody demonstrates beneficial effects on pulmonary inflammation during acute lung injury Pubmed:29456642
Nature Communications Red blood cell-derived semaphorin 7A promotes thrombo-inflammation in myocardial ischemia-reperfusion injury through platelet GPIb Pubmed: 32161256
American Journal of Hematology Semaphorin 7A: A novel marker of disease activity in Gaucher disease Pubmed: 31990411
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPB448Hu01 Recombinant Semaphorin 7A (SEMA7A) Positive Control; Immunogen; SDS-PAGE; WB.
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MAB448Hu22 Monoclonal Antibody to Semaphorin 7A (SEMA7A) WB; IHC; ICC; IP.
SEB448Hu ELISA Kit for Semaphorin 7A (SEMA7A) Enzyme-linked immunosorbent assay for Antigen Detection.
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