ELISA Kit for Semaphorin 3A (SEMA3A)

SEMAL; Hsema-I; Hsema-III; SEMA1; SEMAD; SEMAIII; SemD; Coll-1; Sema Domain,Immunoglobulin Domain(Ig),Transmembrane Domain(TM)and Short Cytoplasmic Domain 3A

Specificity

This assay has high sensitivity and excellent specificity for detection of Semaphorin 3A (SEMA3A).
No significant cross-reactivity or interference between Semaphorin 3A (SEMA3A) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Semaphorin 3A (SEMA3A) and the recovery rates were calculated by comparing the measured value to the expected amount of Semaphorin 3A (SEMA3A) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-90 85
EDTA plasma(n=5) 91-99 95
heparin plasma(n=5) 85-97 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Semaphorin 3A (SEMA3A) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Semaphorin 3A (SEMA3A) 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 3A (SEMA3A) 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) 85-99% 79-95% 78-92% 80-98%
EDTA plasma(n=5) 79-102% 98-105% 92-103% 78-104%
heparin plasma(n=5) 80-96% 98-105% 88-101% 98-105%

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
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
Arthritis Research & Therapy Semaphorin 3A is a marker for disease activity and a potential immunoregulator in systemic lupus erythematosus Biomed: ar3881
journal of invastigative dermatology Cathelicidin LL-37 Induces Semaphorin 3A Expression in Human Epidermal Keratinocytes: Implications for Possible Application to Pruritus PubMed: 26121211
Neurology Neurofilament light chain level is a weak risk factor for the development of MS. pubmed:27521440
Development miR-126-5p promotes retinal endothelial cell survival through SetD5 regulatio in neurons pubmed:29180574
A Preliminary Study of the Effect of Semaphorin 3A and Acitretin on the Proliferation, Migration, and Apoptosis of HaCaT Cells
Life Sciences Curcumin and LOXblock-1 ameliorate ischemia-reperfusion induced inflammation and acute kidney injury by suppressing the semaphorin-plexin pathway. Pubmed: 32603817
ACS NANO Gold Nanoparticles Induce Tumor Vessel Normalization and Impair Metastasis by Inhibiting Endothelial Smad2/3 Signaling Pubmed: 32413258
PLoS One Extract of Scutellaria baicalensis induces semaphorin 3A production in human epidermal keratinocytes 33905439
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